RNA sequencing to compare gene expession in control and PF228-treated hepatic stellate cells

Primary human hepatic stellate cells (HSCs) isolated from healthy donors were purchased from ScienCell Research Laboratories. They were preincubated with or without PF228 and stimulated with TGFbeta1 (5 ng/ml) for 24 hours. Cells were collected for RNA isolation and RNA sequencing. The goal of this study was to identify genes transcriptionally regulated by TGFbeta1 and FAK. There were 4 groups of cells based upon the following treatments: DMSO, DMSO+TGFbeta1, PF228, PF228+TGFbeta1. Each group had 3 repeats, so 12 RNA samples were sent to UMN Genomic Center for RNA sequencing. The RNA samples were converted to Illumina sequencing libraries by using Illumina’s Truseq Stranded mRNA Sample Preparation Kit. Truseq libraries were then subjected to cluster by using Illumina cBot instrument and sequencing was done by using HiSeq2500.