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Fig. 3. RGMc is secreted by differentiating muscle cells. (A) Time course of accumulation of RGMc (arrows) in culture media conditioned by differentiating C2 myoblasts (CM). (B) Detection of RGMc by immunoblotting (arrows) in CM from C2AS12 myoblasts after incubation for 72 hours in differentiation medium (DM) with IGF-I but not PDGF (left panel) and by C3H10T1/2 cells after infection with Ad-MyoD but not Ad-ß-gal, and incubation in DM for 48 hours (right panel). (C) Incubation of CM containing RGMc secreted from C2 myoblasts (T72) with PNGase F (+) or after mock treatment (-). White arrowheads indicate digestion products.