Figure 9. Increased autophagy flux in murine myotubes exposed to ethanol. (A) Representative immunoblots and densitometry for LC3B lipidation in C2C12 myotubes treated with 100 mM ethanol (ETOH), 10 uM chloroquine (CQ), and 100 mM ethanol with 10 μM chloroquine. Lettering above bars denotes differences as follows: (a and b) < 0.001; (a–c) P < 0.05; (a–d) P < 0.01; (b and c) P < 0.01; (c and d) P < 0.01; (b–d) P < 0.01 and identical letters indicate no difference between those groups. (B) Representative fluorescent confocal microscopy images of C2C12 murine myotubes stably transfected with GFP-LC3B and treated with 100 mM ethanol, 10 μM chloroquine and 100 mM ethanol with 10 uM chloroquine. Percentage of cells with at least 5 GFP-LC3B positive and mean number of puncta per cell. (a and b) P < 0.01; (a–c) P < 0.001; (b and c) P < 0.05. Same letters indicate no significant difference. All experiments were done in triplicate and 3 independent experiments were performed.