FIG 3.

Enhanced myogenic differentiation by RSPO2 during the early stage of myogenic differentiation. (A) Differentiating C2C12 cells were treated with the RSPO2 proteins (200 ng/ml) at the different time points indicated. Treatment conditions a to e correspond to image labels and bars a to e in panels B to D. GM, growth medium; DM, differentiation medium; IF, immunofluorescence. (B) Images of MyHC immunofluorescence staining of RSPO2-treated differentiating C2C12 cells. (C) Percentages of MyHC-positive nuclei in total nuclei. (D) Distribution of MyHC-positive cells containing a single nucleus, 2 to 5 nuclei, and more than 5 nuclei in differentiating C2C12 cells treated with RSPO2. Experiments were performed in duplicate, and more than 1,000 total nuclei were counted. Error bars indicate SEM. P values were calculated by Student's t test. **, P < 0.01.