Fig. 4

FTO regulates murine slow-twitch fiber related genes in a NFATC1–YTHDF2- dependent manner. A Quantitative real-time PCR analysis revealed that the mRNA expression of Nfatc1 was significantly decreased in si-Fto treated myoblasts during myogenic differentiation. n = 5 in each case; data are represented as mean ± s.d. B Representative Western blot of NFATC1 in si-Fto treated myoblasts during myogenic differentiation. C Western blot quantification showed that NFATC1 was significantly decreased in si-Fto treated myoblasts. n = 3 in each case; data are represented as mean ± s.d. D The mRNA stability of Nfatc1 was significantly decreased si-Fto treated myoblasts. n = 5 in each case; data are represented as mean ± s.d. E The mRNA expression of FTO was positively correlated with the expression level of NFATC1 in the paraspinal muscle of adolescent idiopathic scoliosis. n = 20 in each case; data are represented as mean. F Quantitative real-time PCR analysis revealed that mRNA expression of Ythdf2 was significantly decreased but Nfatc1, Myh7, and Myh7b were increased in si-Ythdf2 silenced myoblasts with FB23-2 treatment. n = 5 in each case; data are represented as mean ± s.d. G Representative immunohistochemical images of MHC1 (red) in si-Ythdf2 silenced myoblasts with FB23-2 treated during myogenic differentiation. Scale bar, 100 μm. H Immunofluorescence revealed that the MHC1 fusion index was significantly increased in si-Ythdf2 silenced myoblasts with FB23-2 treated. n = 5 in each case; data are represented as mean ± s.d. *P < 0.05 and **P < 0.01