FIGURE 6

SA suppresses autophagy-mediated metastatic processes during starvation or hypoxia. (A) LC3 and p62 expressions in cells co-treated with EBSS or cobalt chloride by western blotting detection. EBSS treatment for 6 h or the hypoxic treatment with cobalt chloride at 200 μM were selected for the following experiments. All values represent the mean ± SD (n = 3, *p < 0.05, **p < 0.01). (B) LC3 and p62 levels in MDA-MB-231 cells with SA (100 μg/ml) alone or combination with either a 6-h treatment of EBSS (Left) or cobalt chloride at 200 μM (Right). Values represent the mean ± SD (n = 3, *p < 0.05, **p < 0.01 vs. Ctrl, ##p < 0.01 vs. EBSS or hypoxia). (C) Representative images of autophagic flux in MDA-MB-231 cells with or without SA (100 μg/ml) during starvation or hypoxic conditions by a LC3-GFP-mRFP reporter. Scale bar: 10 μM. Values represent the mean ± SD (n = 3, *p < 0.05, **p < 0.01 vs. Ctrl, ##p < 0.01 vs. EBSS or hypoxia). (D) Cell counting assays and (E) wound healing assays as well as transwell invasion assays reflected the influences of SA on cell growth or invasiveness under starvation (EBSS) or hypoxia (cobalt chloride). Values represent the mean ± SD (n = 3, *p < 0.05, **p < 0.01 vs. Ctrl, ##p < 0.01 vs. EBSS or hypoxia).