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. 2020 Dec 19;11(5):1227–1245. doi: 10.1016/j.apsb.2020.12.013

Figure 9.

Figure 9

Canonical and non-canonical autophagy both contribute to SIRT3 overexpression induced autophagy. (A) MCF-7 cells transfected with vector or SIRT3 and then treated with different stages of autophagy inhibitors (ULK1 inhibitor SBI-0206965 (10 μmol/L); PI3K inhibitor 3-MA (1 mmol/L); lysosome-mediated proteolysis inhibitors Baf A1 (25 nmol/L) and CQ (5 μmol/L)) to investigate the expression and location of LC3 by immunofluorescence. Cell nucleus were stained with DAPI. Scale bar = 10 μm. ∗∗∗∗P < 0.0001, SIRT3-OE vs. Vector; #P < 0.05, SIRT3-OE+CQ vs. Vector+CQ; ns, no significance. (B) MCF-7 cells transfected with vector or SIRT3 and then treated with different stages of autophagy inhibitors (ULK1 inhibitor SBI-0206965 (10 μmol/L); PI3K inhibitor 3-MA (1 mmol/L); lysosome-mediated proteolysis inhibitors Baf A1 (25 nmol/L) and CQ (5 μmol/L); SBI-0206965+Baf A1; 3-MA+Baf A1 and the expression of SIRT3, SQSTM1/P62 and LC3 were quantified by Western blot. ns, no significance; ∗∗∗∗P < 0.0001 vs. Vector; ####P < 0.0001 vs. SIRT3-OE.