Figure 7. Autophagy is differentially regulated in ischemic BALB/c and BL6 muscle and by BAG3 variants.

A. BL6 and BALB/c mice were subjected to HLI for 3 or 7-days (N≥5 mice per strain, per time-point) and gastrocnemius LC3 mRNA expression (corrected for GAPDH and normalized to BL6 control) was determined by qRT-PCR. *P<0.05 vs. strain-specific control. †P<0.05 vs. strain-specific HLI d3. B, C. LC3II protein abundance was determined in HLI d7 gastrocnemius (Gastroc) by western blotting using GAPDH as a loading control (B) and quantified relative to GAPDH and normalized to non-ischemic BL6 controls (C). *P<0.05 vs. strain-specific control. D. BALB/c mice injected IV with AAVs encoding GFP (N=5), BAG3^Met81 (N=6), or BAG3^Ile81 (N=6) were subjected to modified HLI with collateral vessels left intact, and gastroc LC3 mRNA expression (corrected for GAPDH and normalized to BL6 control) was determined by qRT-PCR. *P<0.05 vs. control. †P<0.05 vs. GFP or BAG3^Met81. E, F. LC3bII protein abundance was determined in Gastroc muscle lysates from BALB/c mice injected IV with AAVs by western blotting using GAPDH as a loading control (E) and quantified relative to GAPDH and normalized BL6 controls (F). In panel (E) all bands are from the same blot and exposure, and the vertical line indicates where the blots were cropped and lanes spliced together for comparison. †P<0.05 vs. GFP or BAG3^Met81. All data are means ± SEM.