Fig. 4. Mettl3 knockdown forces premature myoblast differentiation.

a Mettl3 gene expression levels were assessed via qPCR in C2C12 myoblasts in GM as well 3d and 5d DM (n = 3). ***P < 0.001. b Gene expression levels of Mettl3 were measured via qPCR in GM as well 3d and 5d DM in primary mouse myoblasts (n = 3). ***P < 0.001. c siRNA targeting Mettl3 was applied to C2C12 myoblasts for 2 days in proliferation favoring media and protein levels (immunoblotting) and gene expression (qPCR) levels of Mettl3 demonstrated an efficient knockdown (n = 3). d mRNA was isolated from C2C12 myoblasts after 2 days of siMettl3 or siCON and global m⁶A-modification levels were determined via by LC–MS (n = 6). **P < 0.01. e Total C2C12 myoblast number was counted at the time points indicated after treatment with siMettl3 or siCON (n = 3). *P < 0.05. f Gene expression levels of Pax7, Myod1, and Myog were measured via qPCR after 2 days of siMettl3 or siCON (n = 3). *P < 0.05; ***P < 0.001. g Immunocytochemistry for eMHC in C2C12 cells (left) and quantification (right) after 2 days of siMettl3 or siCON (n = 3). ***P < 0.001.