Fig. 5
From: HMGB1/autophagy pathway mediates the atrophic effect of TGF-β1 in denervated skeletal muscle
Atrophic effect of TGF-β1 in vitro. a, b Quantification of C2C12 myotube diameter by immunofluorescence staining of MHC after treated with a dose gradient of TGF-β1. One-hundred total myotubes were evaluated in each group with the ImageJ software. Scale bar 50 μm. c Representative fluorescent images of C2C12 myotubes expressing GFP-LC3 after TGF-β1 treatment. Scale bar 100 μm. d Quantification of the average number of GFP-LC3 puncta per cell as described in (c). e, f Western blot analysis of HMGB1, LC3B, p62 and MHC protein expression in C2C12 myotubes. Relative grey values analyses were performed. g, h Western blot analysis of HMGB1 in the supernatant after TGF-β1 treatment. i ELISA analysis of HMGB1 in the supernatant after TGF-β1 treatment. The values were obtained from three independent experiments. *P < 0.05 vs control (0 ng/ml)