Genome wide expression analysis of high passage growth arrested BJ fibroblasts maintained or withdrawn from rapamycin maintenance, in the presence or absence of various SMAD2/3 regulating molecules

BJ fibroblasts were grown in the presence of rapamycin until reaching growth arrest at high passages, at which point cells were maintained in or withdrawn from rapamycin, while also being treated with control or SMAD2/3 regulating ligands. RNA was then harvested for RNA-seq Overall design: BJ fibroblasts were grown to growth arrest in the presence of rapamycin maintenance. Cells were then either maintained in rapamycin or withdrawn from rapamycin for two weeks, along with treatment with control or SMAD2/3 regulating ligands Activin A (100ng/ml), TGFB2 (1ng/ml), Inhibin (100ng/ml)+TGFB2 (1ng/ml), or Inhibin (100ng/ml). Experiments, excepting control treatment, were done in duplicates, and each treatment was compared with controls to determine a fold change in transcription for genes.