GSE157717
Next Generation Sequencing Facilitates Quantitative Analysis of Transcriptomes in HeLa cells upon Hippo Components Overexpression or Knockout
Purpose: The goals of this study are to compare and integrate NGS-derived transcriptome when Hippo components are manipulated in HeLa cells. Methods: Total RNAs from three replicates were extracted using TRIzol (Thermo Fisher Scientific) from HeLa cells. RNA quality was assessed using a bioanalyzer (Agilent Technologies). RNA-seq libraries were prepared according to the Illumina TruSeq protocol. Then were pooled for deep sequencing by using Illumina Hiseq Xten (2 × 150) platforms at the Omics Core of Bio-Med Big Data Center, CAS-MPG Partner Institute for Computational Biology, Shanghai, China. Raw read qualities were evaluated with FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/, v0.11.5). Adaptor sequences and read sequences on both ends with Phred quality scores below 30 were trimmed. Trimmed reads were then mapped with the Hisat2 algorithm (version 2.1.0) to target sequences. Gene expression levels were quantified by the software package HTSeq (v0.6.1p1). The list of differentially expressed genes were generated by DESeq2 with FC:1.5,and adjust p-value<0.05.The significant genes were then compared with each other. Results: We finally get 558 genes that have dramatical change in different groups. Conclusions: Our study shows many common genes that can be loyally regulated by Hippo signaling. Overall design: RNA-seq analysis of HeLa cells in which Hippo components are manipulated.