Fig. 1

BAG1 protein sufficient to suppress MYC-driven apoptosis. a U2OS MYC-ER cells depleted of BAG1 via infection with an shRNA-encoding lentivirus directed against the 3’UTR or a luciferase shRNA control. Five days post-infection cells treated with ±100 nM 4-OHT for 48 h. Apoptosis measured by Annexin V-PE plus 7AAD DNA staining and quantified by FACS analysis. b Quantification of three experimental replicates representing average and standard deviation of cumulative early and late apoptosis based on population of Annexin V-PE positive cells. c PARP cleavage and BAG1 knockdown demonstrated by immunoblot (IB). F and C indicate full and cleaved species respectively. d U2OS MYC-ER cells transfected with vectors encoding BAG1 or empty vector, followed by endogenous BAG1 depletion. IB demonstrating efficient knockdown of endogenous BAG1 and retained ectopic BAG1 expression. e Cells from d cultured in ±100 nM 4-OHT for 48 h. Apoptosis measured by Annexin V-PE plus 7AAD DNA staining and quantified by FACS analysis. f Quantification of three experimental replicates representing average and standard deviation of apoptosis based on population of Annexin V-PE positive cells. g PARP cleavage and BAG1 knockdown demonstrated by IB. F and C indicate full and cleaved species respectively. **p < 0.02