Fig. 6From: Hydroxychloroquine synergizes with the PI3K inhibitor BKM120 to exhibit antitumor efficacy independent of autophagyHCQ and BKM120 cooperated to induce DSBs. A Comet assay of MKN-1 cells treated with HCQ and/or BKM120 for 48 h. The %DNA in tails was quantified to represent DNA damage intensity. B Representative images of immunofluorescence staining of γH2AX in MKN-1 cells treated with HCQ and/or BKM120 for 48 h. Cell nuclei were stained with DAPI. Scale bar, 20 μm. The percentage of γH2AX foci positive cells (> 5 foci per cell) is shown in (C). D Western blot analysis of γH2AX in MKN-1 cells treated with BKM120 and/or HCQ in the presence or absence of NAC or Tiron for 48 h. E HR reporter assay was used to detect the effect of the drugs on HR repair efficiency in SKOV-3 and MKN-1 cells. F Western blot analysis of p-ATM, BRCA1, BRCA2 and Rad51 in SKOV-3 and MKN-1 cells treated with each drug alone or in combination for 48 h. Graphs are normalized to the untreated group and shown as the mean ± SD from three independent experiments; **p < 0.01; ***p < 0.001Back to article page