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. 2020 Dec 3;8:601953. doi: 10.3389/fcell.2020.601953

FIGURE 4.

FIGURE 4

Over expression of SIRT2 reduced exosomes and autophagy component release, promoted ASS induced cell apoptosis, inhibited cell invasion. (A,B) A549 and H1299 cells were exposed to ASS (10 dyn/cm2) for indicated times, and then the expression of SIRT1 and SIRT2 were detected by western blot, each experiment was repeated at least three times, **P < 0.01, ***P < 0.001, ****P < 0.0001. (C,D) After overexpression of SIRT2 or treatment with GW4689, A549 cells were exposed to ASS (10 dyn/cm2) for 60 min, quantitative analysis of the released exosome (TSG101, CD63, HSP70) and autophagy component (LC3) markers were carried out by western blot, ****P < 0.0001 compared to control group, $$$$ denote P < 0.0001 compared to ASS (10 dyn/cm2) group, &&&, &&&& denote P < 0.001 and P < 0.0001 compared to OE-NC group. (E,F) Cell apoptosis rate were, respectively, examined by (E) TUNEL assay, and (F) flow cytometry after the cells were stably overexpressed for SIRT2 or pretreated with GW4689 for 60 min. Further, the cells were exposed to ASS for 60 min, *** and **** denote P < 0.001 and P < 0.0001 compared to control group,### denote P < 0.001 compared to ASS (10 dyn/cm2) group, $$, $$$ denote **P < 0.01 and P < 0.001 compared to OE-NC group. (G) After overexpression of SIRT2 or treatment with GW4689, A549 cells were exposed to ASS (10 dyn/cm2) for 60 min, cell invasion was detected by Tranwell assay.