Figure 5. The autophagy inhibitor 3-methyladenine abolished the protective effect of acacetin against H9C2 cell senescence induced by D-galactose. (A) Representative images of SA-β-gal staining for senescent cells in H9C2 cardiac cells treated with D-galactose (D-gal) in the absence or presence of 3 μM acacetin or 10 μM 3-methyladenine (3-MA) with or without 3 μM acacetin for 72 h. (B) Percentage of senescent cells in cells treated as in (A). (C) Western blots and relative level of p21 protein in cells treated as in (A). (D) Western blots and relative level of p53 protein in cells treated as in (A). (n = 4, **P < 0.01 vs. control. ##P < 0.01 vs. D-gal).