Fig. 9. Skeletal muscle dysregulated proteostasis and ethanol-induced phenotype reversed by HMB.(A) Representative immunoblots and densitometry (for the indicated conditions) for puromycin incorporation in C2C12 myotubes that were untreated or treated with 100 mM ethanol with or without 50 µM HMB. (B) Representative immunoblots and densitometry of LC3 lipidation in in differentiated C2C12 myotubes treated with 100 mM ethanol for 6h with and without 50 µM HMB. Chloroquine was used to determine autophagy flux. (C). Representative photomicrographs of C2C12 myotubes treated with and without ethanol and HMB. Mean diameter of at least 100 myotubes in each group. All data mean+SD from at least 3 biological replicates for myotubes and n=4 PF and n=6 mALD mice. *p<0.05; **p<0.01; ***p<0.001. EtOH: ethanol; HMB b-hydroxy-b-methyl butyrate; UnT: untreated controls; PF: pair-fed; mALD: mice with alcoholic liver disease.