MicroRNA Expression Profiling by Bead Array Technology in Human Tumor Cell Lines Treated with Interferon-Alpha-2a
互联网
The gene expression patterns of tumor-derived cell lines differ greatly, as do their responses to antiproliferative effects of interferons (IFNs). The cause of this variation has been under investigation for more than 40 years, but only basic regulatory mechanisms of interferon signaling are understood today. Small regulatory genome encoded RNAs, such as microRNAs, have recently attracted attention in genomic research. New methods to analyze the levels of these regulatory elements are now commercially available, but the power of these techniques is still discussed extensively. Our study was designed to compare two methods for microRNA detection with respect to usefulness in defined cell culture assays. The experimental design assesses variation between the two cell lines and the treatment effects of IFNα.
A hallmark of the therapeutic activity of type I interferons is the induction of antiproliferative activity mediated by the upregulation of several hundred response genes with pleiotropic functions (1 ). These genes can be divided into two major classes based on the kinetic properties of induction (2 ). Primary response genes (PRGs) are upregulated within 24 h after the cytokine signal and the secondary response genes (SRGs) are induced following day 1 when the activity of the PRGs decays. In contrast to SRGs, all PRGs studied to date contain bona fide interferon response elements in the promoter region, which are required for binding of the interferon-stimulated gene factor 3 (ISGF3) complex and for janus kinase/signal transducer of transcription (JAK/STAT)-pathway-mediated signaling.
Expression of PRGs is turned off by proteins termed suppressors of cytokine signaling (SOCS) (3 ). As the nomenclature indicates, this class of polypeptides has the capacity to interfere and silence other cytokine-induced signaling cascades (for review see (4 )). SOCS1 for instance is part of the early inducible PRG cluster and down modulation occurs together with the other genes before onset of SRG expression. It is believed that feedback inhibition of JAK/STAT signaling by SOCS1 represses transcriptome modulation of IFNα signaling (5 ). Regulation of SOCS protein translation by interferon-regulated microRNAs (IRmiRs) would enhance the potential of cytokine fine regulation. It has been reported that miR-19 antagonists lead to higher SOCS1 levels and miR-19 mimics can repress SOCS1 reporter constructs, thus obviously supporting the bioinformatic predictions that SOCS1 is a direct target of miR-19 (6 ). Inhibition of SOCS activity could for instance prolong the duration of cytokine activity, which has obvious clinical implications.
Following the discovery of microRNAs in virtually all higher eukaryotic organisms significant research efforts were initiated to address the function of these catalytic oligonucleotides which are the natural counterparts of synthetic small inhibitory RNAs (siRNAs) used for experimental gene silencing (for review see (7 )). MicroRNAs are positive and negative modulators of the expression of entire gene clusters that contain complementary microRNA recognition sequence motifs in the 3′-UTR. Today, prediction of microRNA target genes by homology-based algorithms is still ambiguous (8 ). The activity of one or several microRNAs could explain suppression of the entire PRG cluster provided that microRNA abundance is regulated by IFNα. Alternatively, microRNA-mediated degradation of transcripts encoding negative regulatory proteins would also abolish PRG expression and restore IFNα responsiveness.
Some recent reports showed that interferon beta (IFNβ) stimulation can boost microRNA levels in cell culture together with inhibition of viral replication (9 ). At this point it is an open question whether this induction is IFNβ specific or a shared feature of all type I interferons. To investigate whether microRNA are also involved in regulation of IFNα response, we used two human-tumor-derived cell lines: the melanoma line ME-15 (10 ) and the hepatoma line HuH7 (11 ). We have chosen these cell lines as models, because we have a good understanding of the IFNα responses at the mRNA and the protein levels in these cell lines. Further we chose to use a melanoma cell line because IFN is also used for treatment of this cancer type. HuH7 is commonly used as a model for testing antiviral effects of IFN in the HCV replicon system. In both models efficient responses to IFNα have been shown at the functional and transcriptional level. IFNα response genes carry response elements in their promoter region and these motifs are responsible for gene expression with similar efficiency in many cell types. Therefore we expected to find a similar regulated set of genes in both lines given that IRmiR genes are regulated by the same mechanism, whereas some constitutively expressed microRNA genes were expected to be cell type specific for functional reasons. We have chosen a DNA-microarray-based technology (Illumina) for the multi-parallel expression analysis of all known human microRNAs (http://microrna.sanger.ac.uk/; Release 10.0: August 2007). This method allowed us to process total RNA as template, allowing the possibility of mRNA gene expression profiling in further experiments. Briefly, annealing of microRNA specific primers combined with enzymatic polyadenylation allows multi-parallel polymerase chain reaction (PCR)-mediated amplification of individual microRNAs. The output of this step is a DNA amplicon library that reflects to a large extent the original stoichiometry of mature microRNAs in a cell or tissue (12 ). PCR amplification is performed with fluorescently labeled primers, which allows quantitative signal detection by conventional confocal laser scanning.
Melanoma cells (ME-15) were cultured in RPMI 1640 with L-Glutamine supplemented with non-essential amino acids and sodium pyruvate (1 mM) and hepatoma (HuH7) cells were cultured in DMEM + GlutaMAX. Both media contained 10% FBS. All cell culture reagents were purchased from Invitrogen (GIBCO®). Roferon (Interferon alpha2a, ROCHE) was diluted in fresh medium to a final concentration of 1,000 U/mL and control cultures were grown without cytokine. Cells were cultured at 37°C in a humidified atmosphere containing 5% CO2 . Total RNA preparation was carried out using TRIZOL (Invitrogen) total RNA extraction using 1/2 volume of 1-bromo-3-chloro-propane (molecular biology grade, SIGMA) as chloroform substitute. For efficient recovery of small RNAs, DNA LoBind tubes (Eppendorf) were used and all centrifugation steps were performed at maximum speed and 4°C in an Eppendorf 5417R centrifuge. Total RNA was precipitated with 2 vol of 2-propanol (Fluka) at −20°C for at least 16 h. The RNA pellet was washed with 75% ethanol (Merck), dried, and dissolved in DEPC-treated water (Ambion). The RNA was quantified with Quant-iT™ RiboGreen® RNA Assay (Invitrogen) as suggested by Illumina.
Starting with 500 ng/sample of total RNA, mature microRNAs were amplified with the Illumina human v1 MicroRNA expression profiling kit containing primers for 743 human microRNAs. The resulting amplicons were hybridized to a 96 sample universal probe capture array and fluorescent signals were detected by confocal laser scanning. All steps were performed according to Illumina’s instructions manual.
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Control |
Interferon-alpha treated |
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4 h |
24 h |
4 h |
24 h |
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ME-15 |
HuH7 |
CHF |
ME-15 |
HuH7 |
CHF |
ME-15 |
HuH7 |
CHF |
ME-15 |
HuH7 |
CHF |
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microRNAs rated higher in HuH7 |
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hsa-miR-122a |
685 |
30,912 |
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~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E227~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E512~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4.6undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E067~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M13~E907~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.5undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F452~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M434~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E284~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M707~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E326~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M530~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E378~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M636~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E028~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.7undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F183~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E763~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E748~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E123~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E945~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E000~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E984~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E129~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E258~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2.8undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F200b~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E934~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E551~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E692~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E633~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4.6undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E705~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E393~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E911~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E855~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4.6undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F143~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E355~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E087~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E775~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E167~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E698~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E688~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E765~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E519~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F624~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E852~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M13~E418~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.7undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E188~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M11~E593~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E479~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M11~E718~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.6undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E872~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E539~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.7undefined~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F99a~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E653~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M20~E232~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.3undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E887~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M20~E645~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E064~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M18~E019~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M0.9undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E870~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M19~E506~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.2undefined~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F27b~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E067~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M22~E544~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.2undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M11~E924~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M23~E914~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M11~E564~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M23~E044~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M0.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E182~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M24~E515~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1.0undefined~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~MmicroRNAs_rated_higher_in_ME~F15~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F146a~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M36~E976~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E721~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.3undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M33~E410~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M16~E103~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M35~E166~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E137~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M35~E688~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M19~E962~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−0.7undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F422b~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M19~E264~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E759~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.4undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M19~E962~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E832~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M20~E641~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E462~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M17~E453~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E152~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.4undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F149~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E868~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E246~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.6undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E195~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E030~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E061~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E743~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E477~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E418~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.2undefined~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F510~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E051~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M368~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E489~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M396~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E435~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M366~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E053~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M384~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.7undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F30a~F5p~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E664~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E717~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.8undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E223~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E700~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E686~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E477~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E466~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E538~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−1.3undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~MHS~S98~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E274~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E065~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.0undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E225~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M859~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E445~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M965~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E850~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M750~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.8undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F340~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E583~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E443~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.1undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E218~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E220~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E664~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E061~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E078~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M933~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.3undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~MHS~S182.1~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E965~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M615~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.undefined* |
1,687 |
606 |
−1.7undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E903~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M562~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E844~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M534~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.4undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F378~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E619~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E371~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.3undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E975~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E364~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.6undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E994~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E213~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.1undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E188~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E237~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.3undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~MHS~S305~Sb~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E105~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E207~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.4undefined |
3,853 |
833 |
−3.6undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E278~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M977~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E688~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E076~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.4undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F505~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E312~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M343~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E070~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M342~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E050~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M360~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E030~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M362~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.6undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F10a~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E570~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M350~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.4undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E592~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M350~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E828~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M360~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E880~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M352~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.3undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F10b~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E695~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M378~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.4undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E929~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M371~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E564~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M411~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.6undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E297~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M382~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~Flet~F7g~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E720~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E302~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.6undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M16~E046~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E077~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M13~E792~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E854~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E435~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E568~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.0undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F584~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M11~E643~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E261~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.1undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E717~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E873~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E394~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E912~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E616~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E674~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.5undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~Flet~F7i~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M11~E701~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E255~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.1undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M16~E410~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E707~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E319~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E251~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E718~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E563~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.1undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F330~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E029~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E653~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E684~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E670~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E432~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E397~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E773~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E380~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.6undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~MHS~S307~Sb~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E453~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M444~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E188~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M443~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−8.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E669~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M407~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−8.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E318~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M448~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.4undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F34c~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E292~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M412~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.5undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E303~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M426~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E524~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M381~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−8.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E403~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M404~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.9undefined~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F133a~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E270~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E594~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E942~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E662~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−3.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E488~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E435~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−4.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E495~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E783~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−2.6undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F361~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E234~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M828~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E641~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M755~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−9.1undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E356~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M875~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M7~E904~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M706~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.1undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F508~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M2~E552~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M325~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.8undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E280~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M376~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E965~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M357~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E013~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M336~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.9undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~Fundefined |
8,372 |
1,039 |
−7.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E114~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M940~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−9.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E027~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M1~E263~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E948~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M843~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.8undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F31~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E967~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M661~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−8.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E742~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M720~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−6.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E619~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M657~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−9.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E978~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M728~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F506~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E855~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M625~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−8.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E125~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M713~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E916~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M665~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.9undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E602~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M675~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.3undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F211~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E147~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M476~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−9.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E218~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M490~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−5.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M3~E833~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M471~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.1undefined~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E575~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M464~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−11.0undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F296~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E194~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M570~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−9.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M5~E913~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M504~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M6~E441~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M557~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E369~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M532~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F598~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E445~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M401~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E475~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M386~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E934~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M424~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.6undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M4~E127~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M367~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F199b~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E081~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M775~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.7undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E478~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M916~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−9.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E629~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M865~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−10.1undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E059~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M911~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−7.8undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~Flet~F7b~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M8~E236~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M539~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−14.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E563~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M475~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−25.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E150~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M500~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−23.3undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E309~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M452~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−26.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F509~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E413~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M536~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−18.4undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M10~E346~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M512~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−19.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E966~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M548~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−17.2undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M9~E311~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M529~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−16.6undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F9~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E587~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M422~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−28.8undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E870~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M447~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−34.5undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M15~E502~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M418~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−36.0undefined*~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M16~E402~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M429~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M−37.2undefined*~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~Ktr~M~Ktd~M~1~Kp~Mhsa~FmiR~F514~K~Hp~M~1~K~Htd~M~1~Ktd~M~1~Kp~M12~E065~K~Hp~M~1~K~Htd~M~1~K~Htr~M~1~K~Htbody~M~1~K~Htable~M~1~1~Kp~M364~K~Hp~M~1~1~1~Kp~M−32.1undefined*~K~Hp~M~1~1~1~Kp~M13~E112~K~Hp~M~1~1~1~Kp~M358~K~Hp~M~1~1~1~Kp~M−35.6undefined*~K~Hp~M~1~1~1~Kp~M12~E201~K~Hp~M~1~1~1~Kp~M315~K~Hp~M~1~1~1~Kp~M−37.7undefined*~K~Hp~M~1~1~1~Kp~M12~E355~K~Hp~M~1~1~1~Kp~M355~K~Hp~M~1~1~1~Kp~M−33.8undefined*~K~Hp~M~1~1~1~1~Kp~MHybdridization_controls~K~Hp~M~1~1~1~1~Kp~Marray~Shyb~Scon4~K~Hp~M~1~1~1~Kp~M6~E404~K~Hp~M~1~1~1~Kp~M7~E110~K~Hp~M~1~1~1~Kp~M0.11~K~Hp~M~1~1~1~Kp~M7~E724~K~Hp~M~1~1~1~Kp~M6~E684~K~Hp~M~1~1~1~Kp~M−0.16 7,031 6,739 −0.04 6,964 6,746 −0.03 array_hyb_con3 6,923 7,681 0.11 7,833 7,368 −0.06 7,422 7,054 −0.05 6,906 7,365 0.07 array_hyb_con1 10,779 11,868 0.10 11,812 11,728 −0.0.01 11,436 11,382 0.00 11,059 11,522 0.04 array_hyb_con2 10,258 11,145 0.09 11,382 10,829 −0.05 11,075 10,650 −0.04 9,997 10,283 0.03 List of calculated relative fluorescent levels (without background correction) and calculated differences (CHF) between hepatoma (HuH7) and melanoma (ME-15) cells. A linear model with ME-15 and HuH7 as separate factors was used to estimate cell line differences in microRNA expression. The genes with the highest significance (and log factor change >0.5) are listed in the upper section and the bottom shows data for the hybridization controls as reference. Change factors show the cell line differences of the retransformed mean to the linear scale. p
values (limma t
test) were adjusted by false discovery rate correction. Significance codes are defined by the intervals: ‘**’ < 0.001 ≤ ‘*’ < 0.01 ≤ ‘’ < 0.05
Table 2
Modulation of microRNA expression by IFNα―4 and 24 h after stimulation
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