Fig. 7. Ethanol-induced increased in free radical generation is reversed by HMB. Studies in differentiated C2C12 myotubes treated with/without 100mM ethanol with/without 50 mM HMB for 6h. (A). Representative flow cytometry of myotubes gated for MitoSox fluorescence and percentage of MitoSox fluorescent cells. (B). Representative immunoblots of carbonylated proteins and densitometry, and TBARS in myotubes treated with/without 100mM ethanol with/without 50 mM HMB for 6h. (C) Representative immunoblots and densitometry of carbonylated proteins , and tissue concentration of TBARS in gastrocnemius muscle from mALD and PF mice treated with/without HMB. (D) Representative immunoblots and densitometry of voltage dependent anion channel (VDAC) and citrate synthase (CS) expression in myotubes as measures of mitochondrial mass. (E). Representative immunoblots of VDAC and CS in gastrocnemius muscle from PF mice and mALD and PF mice treated with and without HMB. All data expressed as mean±SD from at least three biological replicates for experiments in C2C12 myotubes and n=4 PF and n=6 mice. *p<0.05; **p<0.01; ***p<0.001. EtOH: ethanol, TBARS: thiobarbituric acid reactive substances; UnT: untreated controls; PF: pair-fed; mALD: mouse model of ALD.