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Fig. 1. Induction of RGMc mRNA and protein expression during myogenic differentiation. (A) Time course of accumulation of RGMc mRNA and mRNAs for myogenin, myosin heavy chain (MHC) and S17 over 96 hours of differentiation of C2 myoblasts, as measured by RT-PCR. Relative increases in mRNA levels compared with T₀ are listed below each time point. (B) Time course of accumulation of RGMc, myogenin and -tubulin over 96 hours of differentiation of C2 myoblasts, as measured by immunoblotting using whole-cell protein extracts. The arrows indicate different immunoreactive RGMc polypeptides. (C) Detection of RGMc in whole-cell protein extracts from C3H10T1/2 cells infected with Ad-ß-gal or Ad-MyoD, and incubated in differentiation medium (DM) for 48 hours. Arrows indicate the immunoreactive RGMc proteins. (D) One major RGMc mRNA species is synthesized in differentiating mouse muscle cells. The upper panel shows a schematic of the mouse RGMc gene. Exons are depicted as boxes, with predicted coding regions in black and introns are indicated by dotted lines. The relative positions of primers and predicted product sizes are displayed below the gene. The lower panel shows results of RT-PCR experiments, using exon-specific primers and RNA from differentiating muscle cells (C2AS12 and C2 myoblasts, 72 hours in DM, Ad-MyoD infected C3H10T1/2 cells, 48 hours). Positions of molecular mass markers are indicated to the left of blots.

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