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. 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 is to identify genes transcriptionally regulated by TGF-beta1 and FAK. There were 4 cell groups in the experiments: DMSO, DMSO+TGFbeta1, PF228, PF228+TGFbeta1. Overall design: There were 4 cell groups in the experiments: DMSO, DMSO+TGFbeta1, PF228, PF228+TGFbeta1, and each group had 3 repeats. So 12 RNA samples were sent to UMN genomic Center for RNA sequencing. 12 RNA samples were converted to Illumina sequencing libraries using Illumina's Truseq Stranded mRNA Sample Preparation Kit. Truseq libraries were then subjected to cluster using Illumina cBot instrument and sequencing using HiSeq2500

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Wang Y, Tu K, Liu D, Guo L, Chen Y, Li Q, Maiers JL, Liu Z, Shah VH, Dou C, Tschumperlin D, Voneschen L, Yang R, Kang N