Figure 3.

Receptor for advanced glycation end‐products (RAGE) activity is critical for Lewis lung carcinoma (LLC)‐induced muscle wasting. (A–D) Gastrocnemius muscles from LLC‐wild type (WT) and LLC‐Ager ^−/− mice were analysed compared with internal control mice (Ctrl) at the indicated dpi. (A) myoblast determination protein 1 (MyoD), myogenin and the adult fast [myosin heavy chain (MyHC)‐II] and slow (MyHC‐I) myosin heavy chain isoforms were detected by western blot (WB). Reported are the relative densities. (B) Expression of developmental MyHC (dMyHC) at 25 dpi was analysed by western blot (WB). α‐Actinin or GAPDH were used for loading control (A,B). (C) Levels of Fbxo32, Trim63 and Myog were analysed by real‐time PCR. (D) Myogenin (red) and atrogin‐1 (green) were detected by immunofluorescence (IF). 4′,6‐diamidino‐2‐phenylindole (DAPI) (blue) was used to stain nuclei. See also Figure  S5 . Reported are high‐magnification insets with myofibers defined by dashed lines. Results are means ± standard error of the mean. Statistical analysis was conducted using the two‐tailed t‐test. ^* P < 0.05, ^** P < 0.01, and *** P<0.001, significantly different from internal control mice. ^# P < 0.05, ^## P < 0.01 and ^### P < 0.001 significantly different. ^$ P < 0.05 and ^$$ P < 0.01 LLC‐Ager ^−/− vs. LLC‐WT mice significantly different (C). Scale bars in (D), 25 μm.