Comparative miRNA expression profiling analysis of RNA-seq data of hESCs WA09 (H9) cells (CONTROL) and these cells treated with 20 uM 24 hours pan-E2F inhibitor treatment (iE2F-20uM)

To investigate those miRNAs regulated by E2Fs transcription factors in hESCs we treated WA09 (H9) hESCs with a 20 mM 24 hours pan-E2F inhibitor (HLM006474).We then performed miRNA expression profiling analysis using data obtained from RNA-seq of three biological replicates of H9 vehicle control and three biological replicates of pan-E2F inhibitor treated cellsComparative miRNA expression expression profiling analysis of RNA-seq data of hESCs WA09 (H9) cells (CONTROL) and these cells treated with 20 mM 24 hours pan-E2F inhibitor treatment (iE2F-20mM).hPSCs were cultured on Geltrex (Thermo Scientific) coated dishes in combination with a fully defined mTeSR1 medium (Stem Cell Technologies) in humidified atmosphere with 5% CO2 at 37°C.hESCs WA09 (H9) cells were treated with 20 mM 24 hours pan-E2F inhibitor HLM006474 (Sigma).Total RNA was extracted with mirVanaTM miRNA Isolation Kit (Ambion, Thermo Fisher Scientific) following the manufacturer’s protocols.Small RNA libraries for small RNA-seq were prepared using the TruSeq small RNA Library Prep Kit following manufacturer’s protocols.