Fig. 5

Concurrent treatment with HCQ and BKM120 induced ROS accumulation and apoptosis. A Determination of ROS production in MKN-1 cells treated with BKM120 and/or HCQ for 48 h. B Western blot analysis of NRF2 in SKOV-3 and MKN-1 cells treated with BKM120 and/or HCQ for 48 h. C The mRNA levels of Nqo1 and Hmox1 in SKOV-3 cells treated with BKM120 and/or HCQ for 48 h were determined by qRT-PCR. D Cell viability of SKOV-3 and MKN-1 cells after treatment with BKM120 and HCQ in combination with NAC or z-VAD for 72 h. E Colony-forming ability of SKOV-3 and MKN-1 cells treated with BKM120 and HCQ in the presence or absence of NAC or z-VAD. Colony quantification is graphed in (F). G SKOV-3 and MKN-1 cells were treated with BKM120 and HCQ in combination with NAC or z-VAD for 48 h and then subjected to Annexin V-FITC/PI staining and flow cytometric analysis of apoptosis. H Western blot analysis of Caspase-3 and PARP in SKOV-3 and MKN-1 cells treated with BKM120 or/and HCQ in the presence or absence of NAC for 48 h. Graphs are normalized to 100% per treatment and shown as the mean ± SD from three independent experiments; *p < 0.05; **p < 0.01; ***p < 0.001