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. 2016 Jul 1;7(30):48501–48516. doi: 10.18632/oncotarget.10363

Figure 3. Inhibition of autophagy by CQ and si-BECN1 enhanced the cytotoxicity of gemcitabine in ER-negative MDA-MB-231 cells.

Figure 3

A. The proliferation of MDA-MB-231 cells exposed to gemcitabine alone (0.16-2.5 μg/ml) or gemcitabine + CQ (5 μM, added before gemcitabine treatment for 1 h) for 12 h and 48 h was detected by MTT assay. The results were expressed as the relative percentage of each group compared with the control. The results (mean ± SE) were from three independent experiments. B, C. Cell viability of MDA-MB-231 cells was measured with AV-FITC/PI through FACS after treated with gemcitabine (1 μg/ml) for 0, 12, 24, 48 h or gemcitabine plus CQ for 12 h and 48 h. Scatter plots of Annexin V-FITC and PI staining were shown. The results of statistical analysis were showed in the bar graphs. D. The levels of Beclin 1 protein were detected by western blotting after treatment with scramble siRNA or si-BECN1 for 0 and 48 h in MDA-MB-231 cells. E. Cells were treated with scramble siRNA or si-BECN1 for 48 h, incubated with or without gemcitabine (1 μg/ml) for 12 h and 48 h. The proliferation was measured by MTT assay and the results were expressed as the relative percentage of each group compared with the control. The results (mean ± SE) were from three independent experiments. F. MDA-MB-231 cells were treated with or without scramble siRNA or si-BECN1 for 48 h, followed by treatment of 1 μg/ml gemcitabine for 12 h or 48 h, and the cell viability was determined by AV-FITC/PI staining by FACS. The results (mean ± SE) were from three independent experiments. *, P<0.05; **, P<0.01;***, P<0.001.