Recombinant BMP3B (R&D Systems) was dissolved in a serum-free medium. The effect of BMP3B on human myotubes was examined similarly to that of the conditioned culture. Recombinant GDF8/myostatin (R&D Systems) and BMP4 (R&D Systems) were used as positive controls for Smad2 and Smad1/-5/-8 signaling pathways.

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