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. 2015 Mar 18;5:8. doi: 10.1186/s13395-015-0032-z

Figure 2.

Figure 2

Retinoic acid partially rescues the inhibition of myogenesis induced by TGFβ. (A) Representative images of C2C12 myoblasts differentiated in low serum conditions in the continual presence of RA or TGFβ as indicated or with vehicle (Veh) for 96 h and immunostained for myosin heavy chain (MyHC) expression. DAPI is used to reveal nuclei (blue). (B) Differentiation index (#myonuclei/#total nuclei) of C2C12 cells cultured and treated as in (A). Means indicated with the same letter are not statistically different from one another. Means with different letters are significantly different from one another with a P < 0.05, n = 3. (C) Fusion index (#myonuclei/#myotubes) of C2C12 cells cultured and treated as in (A). Calculations include mononucleated myosin heavy chain positive cells. Means with different letters are significantly different from one another with a P < 0.05, n = 3. (D) RT-qPCR analysis of Smad3, Smad2, Pax7, and myogenic marker expression in C2C12 myoblasts treated and differentiated as in (A). Means with different letters are significantly different from one another with a P < 0.05, n = 3. (E) Western analysis of phosphorylated Smad2/3 (pSmad2/3) expression in C2C12 cells in growth conditions. Quantification by densitometry is represented as pSmad2 or pSmad3 relative to actin, respectively, n = 3, **P < 0.01, NS = not significant. Protein levels for Smad2 and Smad3 are shown.