Fig. 2.

Autophagy is increased in MsrA-/- VSMC. (A) Immunoblot of autophagy marker LC3 in MsrA-/- and WT VSMC with or without bafilomycin a1 (Baf) for indicated times. Upper band represents full-length LC3-I and lower band active, cleaved LC3-II. Positive control for autophagy: treatment with serum-free media (SF) for 2 h. (B) Quantification of LC3-II in MsrA-/- and WT VSMC relative to LC3-II in WT VSMC at 0 h; data were normalized to GAPDH (n = 4–5 biological replicates). * p < 0.05 versus WT by 2-way ANOVA. (C) Immunoblots for p62 and GAPDH in in MsrA-/- and WT VSMC with or without bafilomycin a1 (Baf) for 24 h. (D) Data in (C) were normalized to GAPDH and expressed relative to p62 in WT VSMC at 0 h. n = 8 biological replicates * p < 0.05 versus WT at untreated and # p < 0.05 vs MsrA-/- untreated by 1-way ANOVA. (E) MsrA-/- and WT VSMC were transduced with an adenovirus expressing LC3-GFP for 72 h. To some samples, bafilomycin was added for 24 h. Nuclei: blue (TOPRO). Scale bars 20 µm. (F) GFP puncta were quantified per cell, adjusted for cell area and expressed per 1000 µm². n = 5 biological replicates. * p < 0.05 versus WT untreated # p < 0.05 vs MsrA-/- untreated by 1-way ANOVA.