BET inhibition induces apoptosis by modulating PTEN/AKT. (a) Flow-cytometry assays by Annexin V/7AAD staining show cell death rates in OTX015-exposed SKOV3 cells (1 μM and 3 μM) and DMSO controls. Representative dot plots with mean values of two independent experiments showing the percentage of viable cells (quadrant LL, Annexin V−/7AAD−), early apoptotic cells (quadrant LR, Annexin V+/7AAD−), late apoptotic cells (quadrant UR, Annexin V+/7AAD+) and necrotic cells (quadrant UL, Annexin V−/7AAD+). (b) WB analysis of the apoptotic markers cleaved PARP, BCL2 and Caspase-3 in OTX015-exposed SKOV3 cells (0–1–3 μM) for 72 h. Protein analysis showing the strong PTEN increase and the downregulation of phospho (p)-AKT levels in OTX015-exposed SKOV3 (0–1–3 μM) for 72 h. Tubulin served as loading control and representative blot was shown. (c) WB experiments showing PTEN and cleaved PARP expression in SKOV3 cells transfected with PTEN-vector or pcDNA3.1 empty vector and treated with or without 1 μM OTX015 for 72 h. Tubulin served as loading control and representative blot was shown. The original Western Blot images can be found in Figure S4.