FIGURE 5

FIGURE 5

SA inhibits autophagic flux in metastatic cells. (A) LC3-II and p62 expressions in MDA-MB-231 and BT-549 cells in exposure to different concentrations (0–200 μg/ml) of SA for 48 h. (B) Representative electron micrographs of the cells with SA (100 μg/ml) for 48 h. The yellow arrows represented autophagosomes, whereas the red arrows represented autolysosomes. (C,D) Fluorescence photographs of MDA-MB-231 and BT-549 cells transfected with an LC3-GFP-mRFP reporter after SA treatment (0–200 μg/ml) for 48 h. Scale bar: 10 μM. (E) Representative bands of LC3-II and p62 in SA-treated MDA-MB-231 cells in addition to 3-MA (10 mM), wortmannin (Wort, 2 μM), CQ (30 μM), or bafilomycin A1 (Baf1, 100 nM) for 48 h. (F,G) The fluorescence photographs of SA-treated cells labeled with LysoRed or DQ-BSA. Scale bar: 20 μM. All values represent the mean ± SD (n = 3, *p < 0.05, **p < 0.01).