Fig. 5

Lmcd1 induces protein synthesis and myotube hypertrophy in mouse primary myotubes. a Representative microscopy image of differentiated mouse primary myotubes 2 days after transduction with GFP (control) or LMCD1 adenovirus (× 200; n = 8; scale bar = 100 μm) and corresponding myotube diameter measurements. b Total protein/genomic DNA ratio determined in myotubes transduced as in (a) (n = 8). c Protein synthesis determination by puromycin incorporation in differentiated primary mouse myotubes transduced as in (a), and respective quantification (n = 8). d qRT-PCR determination of peroxisome proliferator-activated receptor gamma coactivator-1alpha1 (Pgc-1α1), Pgc-1α4, myostatin, insulin-like growth factor 1 (Igf-1), atrogin, and muscle RING-finger protein-1 (Murf1; n = 8) in differentiated primary mouse myotubes transduced as in (a). e qRT-PCR of myosin heavy chain (Myhc) I, IIa, IIb, and IIx (n = 8) in differentiated primary mouse myotubes transduced as in (a). f Western blot of LMCD1 protein expression in differentiated primary mouse myotubes after 2 days transduction of scrambled shRNA or shLmcd1 adenovirus (n = 8). g Representative microscopy image of differentiated mouse primary myotubes (× 200; n = 8; scale bar = 100 μm) transduced as in (f). h Total protein/genomic DNA ratio in differentiated primary mouse myotubes transduced as in (f) (n = 8). j qRT-PCR for Lmcd1, Pgc-1α1, Pgc-1α4, myostatin, Igf-1, atrogin, and Murf1 (n = 8) in differentiated primary mouse myotubes transduced as in (f). i qRT-PCR of MyhcI, IIa, IIb, and IIx (n = 8) in differentiated primary mouse myotubes transduced as in (f). Data is shown as mean ± SD and *p < 0.05; **p < 0.01; ***p < 0.001