Fig. 3.

Muscle proteolysis and autophagy are hindered by JQ1 administration. a Representative immunoblot showing fast myosin heavy chain (MyHC) protein expression in TA whole extracts from the six animal groups. Upper panel shows quantification of immunoblot bands from four animals per group. Data represent means ± SD. b TA extracts from control and C26-tumor-bearing mice (animals per group: n = 3) were used in an in vitro degradation assay to reveal fast MyHC degradation in control vs. tumor-bearing mice treated with vehicle or JQ1 (−/+). c, d Total RNA was extracted from TA muscles from control and C26-tumor-bearing mice (animals per group: n = 10) treated with vehicle or JQ1 (−/+) and expression levels of MuRF1, MAFbx/Atrogin-1 were measured by quantitative RT-PCR. Data represent means ± SD. e Representative western blot of MAFbx/Atrogin-1 on TA whole extracts (animals per group: n = 4). Bands quantifications are shown in the upper panel. Data represent means ± SD. f–h Quantitative RT-PCR of autophagy genes (Bnip3, GABARAPL1, Cathepsin L) from control and C26-tumor-bearing mice. Ten animals were used for each experimental group. Data represent means ± SD. i, j Representative western blot for LC3 and p-Ulk1 in TA extracts of control and C26-tumor-bearing mice (animals per group: n = 4). Upper panel: quantification of normalized band intensity. Data represent means ± SD. Statistical analysis was performed by using one-way ANOVA followed by Tukey’s post hoc test. *p < 0.05; **p < 0.01; ***p < 0.001. “a” indicates statistical significance compared to control; “b” indicates statistical significance compared to C26 + vehicle; “c” indicates statistical significance compared to C26 + (−)-JQ1 20 mg/kg/day; “d” indicates statistical significance compared to C26 + (−)-JQ1 50 mg/kg/day