Research Paper Volume 15, Issue 21 pp 12618—12632

Figure 1. HMGB1 inhibits Pax-7 synthesis in skeletal muscle myoblasts. After exposing C2C12 cells to HMGB1 for 24 h, the expression of Pax-7 mRNA and protein was examined by (A) qRT-PCR (n = 3) and (B) western blotting assay (n = 3). C2C12 cells were cultured in growth medium (GM) and differentiation medium (DM) for three days with or without HMGB1 (10 ng/mL) for 24 h, after which Pax-7 expression was examined using (C) qRT-PCR (n = 3) and (D) western blotting assay (n = 3). (E) Immunofluorescence staining showing the expression of Pax-7 and desmin in C2C12 cells cultured in GM and DM after HMGB1 treatment (n = 3). Pax-7 is indicated by red coloring, desmin is indicated by green, and DAPI is shown in blue. Scale bars = 50 μm. (F) Number of positively stained cells were quantified by ImageJ software (n = 3). β-Actin was used as the loading control. All data are presented as the mean ± SD of triplicate experiments. ^*p < 0.05 compared with control group; ^#p < 0.05 compared with HMGB1-treated group. Abbreviations: GM: growth medium; DM: differentiation medium.