Figure 6: Prox1 regulates myoblast differentiation.

(a) Analysis of Prox1 mRNA and protein during C2C12 myoblast differentiation (indicated by the increase in MyoD1 mRNA). (b) Prox1 protein and mRNA in human myoblasts before and after differentiation. Primary myoblast lines from two different individuals were analysed. (c–f) Analysis of myoblast proliferation and differentiation-related CyclinD1, Myf5 and MyoD mRNAs in shProx1 and shScr transduced primary human myoblasts before (c) and after differentiation for 7d (d), as well as in mouse C2C12 myoblasts before (e) and after (f) differentiation. (g) Myosin staining of C2C12 cells after 7d of differentiation. Note that Prox1 silencing completely blocked myotube development, as only occasional cells expressed myosin and these cells were negative for shProx1-GFP. Data is presented as mean±s.e.m., n=3+3 biological replicates in a–g (all experiments were repeated three times). One-way analysis of variance with repeated measures followed by Tukey’s posthoc test and Student’s two-tailed unpaired t-test, *P<0.05, **P<0.01, ***P<0.001. Scale bars, 50 μm.