Genome wide mapping of the effect of SMAD2/3 regulating treatments upon the epigenomic landscape in high passage growth arrested cells being withdrawn from rapamycin

BJ Fibroblasts were maintained in rapamycin and were passaged until reaching growth arrest at very high passages. These cells were then maintained in or withdrawn from rapamycin, during which time they were given either control treatments or treatments with factors regulating SMAD2/3 activity. BJ Fibroblasts were maintained in rapamycin and were passaged until reaching growth arrest at very high passages (>PD75). These cells were then maintained in or withdrawn from rapamycin for 2 weeks, during which time they were given either treated with control or the SMAD2/3 regulating ligands Activin A (100ng/ml), TGFB2 (1ng/ml), Inhibin (100ng/ml)+TGFB2 (1ng/ml), or Inhibin (100ng/ml). Cells were then harvested for ChIP-seq of enhancer marking histone modifications H3K27ac or H3K4me2, or the transcription factors p65, SMAD2/3, or NFI. ChIP-seq tag density was then compared between rapamycin withdrawal and maintained samples, as well as between treatment and control samples.