RNA-seq analysis of the effect of loss of Tp53, NF1 or CBS in telomerase-reverse transcriptase (TERT)-immortalized human BJ fibroblasts (BJ-TERT) overexpressing myristoylated AKT1

Overexpression myristoylated AKT1 in human BJ-TERT fibroblasts induces senescence (AIS). We depleted CBS, Tp53 or NF1 gene by shRNA in AIS cells and performed RNA-seq analysis to investigate the regulatory effect of these genes in AIS maintenance. Overall design: BJ-TERT cells were transduced with pBabe-puro empty vector (EV) or pBabe-puro-myrAKT1 to induce senescence. On day 6 post-transduction myrAKT1 cells were transduced with either pGIPZ-shRNA against CBS, Tp53 and NF1, or the control vector pGIPZ-NTC, pBabe-EV cells transduced with pGIPZ-shRNA non-targeting control (NTC). Total RNA was extracted on day 14 post-transduction for RNA-seq. There are five samples including pBabe-NTC, AKT1-NTC, AKT-shTP53, AKT-shNF1, AKT-shCBS with three replicates.