Fig. 4

HCQ enhanced the antitumor activity of BKM120 on Atg5-deficient cells. A SKOV-3 and MKN-1 cells were transfected with non-targeting or Atg5 siRNA, and then treated with HCQ and various concentrations of BKM120 for 72 h. Cell viability was determined by PrestoBlue assay. B SKOV-3 and MKN-1 cells were transfected with or without Atg5 siRNA and were then treated with BKM120 and/or HCQ for 72 h and allowed to recover in fresh media for 7–10 days. The colony-forming ability of the cells was determined. Colony quantification is graphed. C SKOV-3 and MKN-1 cells were transfected with or without Atg5 siRNA and then treated with HCQ and/or BKM120 for 48 h. The treated cells were subjected to Annexin V-FITC/PI staining and flow cytometric analysis of apoptosis. The apoptotic cell population is quantified. D Western blot analysis of Caspase-3 and PARP in SKOV-3 and MKN-1 cells transfected with Atg5 siRNA and then treated with HCQ and BKM120 for 48 h. E DU145 cells were treated with BKM120 alone or in combination with HCQ for 72 h and subjected to cell viability assays. Graphs are normalized to 100% per treatment and shown as the mean ± SD from three independent experiments; ns: p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001