Additional file 1 of Hydroxychloroquine synergizes with the PI3K inhibitor BKM120 to exhibit antitumor efficacy independent of autophagy

Additional file 1: Figure S1. Inhibition of PI3K suppressed cell proliferation and induced apoptosis in tumor cells. (A) The three tumor cell lines were treated with different concentrations of BKM120 for 48 h. Cell viability was determined by PrestoBlue assay. (B) The cells were treated with indicated concentrations of BKM120 for 48 h and subjected to Annexin V-FITC/PI staining and flow cytometric analysis of apoptosis. (C) FACS quantification of total apoptotic cell population including Annexin V+/propidium iodide−early apoptotic cells and Annexin V+/propidium iodide+late apoptotic cells. (D) Western blot analysis of Caspase-3, PARP, LC3B and p62 after treated with BKM120 for 48 h. (E) Western blot analysis of p-Akt, Akt and p-mTOR after treatment with BKM120 for 48 h. (F) Representative images of autophagosome puncta in three cell lines treated with BKM120 or/and HCQ for 24 h. Scale bars, 20 μm. (G) Quantification of autophagosome puncta positive cells (> 10 puncta per cell). All data are mean ± SD from three independent experiments; *p < 0.05; **p < 0.01; ***p < 0.001.