FIG. 1.

Statins induce cardiac autophagy. (A) Time course ranging from 6 to 30 h examining statin-mediated autophagy induction via LC3-I to LC3-II conversion by immunoblot. (B) HL-1 cells infected with adenovirus for LC3-GFP and treated with vehicle or statin, showing autophagic puncta formation by fluorescence microscopy. (C) Quantitation of autophagy induction by statin treatment. Cells with >20 puncta were scored positive for autophagy. n=3 plates per group; 50 cells were scored for each n. (D) Autophagic flux was examined via use of 100 nM bafilomycin A1 for 2 h immediately before 24 h statin (or vehicle DMSO) treatment of cells. p62/SQSTM1 and LC3 were monitored. (E) Quantitation of results shown in (D) (DMSO, solid bars; and simvastatin, hashed bars). n=4 per group; experiments were performed at least two times. *p-value<0.05. (F) Representative western blot for LC3 from hearts of mice given simvastatin (20 mg/kg) i.p. and sacrificed 4 h after treatment. n=3 mice per group; experiments were performed twice with similar results. *p-value<0.05. DMSO, dimethyl sulfoxide; GFP, green fluorescent protein; i.p., intraperitoneal; LC3, light chain 3.