Fig. 6.

YBX2 and SMO inhibit adipogenesis and promote myogenesis in m⁵C-dependent manner. A Western blotting of NSUN2 and YBX2 expression in PFs transfected with shCTL, shNSUN2, vector, NSUN2-WT or NSUN2-MUT adenovirus. ACTB was used as a loading control. The relative protein expression was quantified by densitometry and normalized to ACTB. All data are shown as means ± S.D. B m⁵C-RIP-qPCR analysis of YBX2 in PFs transfected with shCTL, shNSUN2, Vector or NSUN2-WT adenovirus. Error bars, means ± S.D., n = 3. C Western blotting analysis in control or YBX2 knockdown PFs transfected with NSUN2-WT or NSUN2-MUT adenovirus. ACTB was used as a protein loading control. The relative protein expression was quantified by densitometry and normalized to ACTB. All data are shown as means ± S.D. D Oil Red O staining analysis in control or YBX2 knockdown PFs transfected with NSUN2- WT or NSUN2-MUT adenovirus. Scale bar: 100 μm. E qPCR analysis in control or YBX2 knockdown PFs transfected with NSUN2-WT or NSUN2-MUT adenovirus. 18S rRNA served as an internal RNA control. Error bars represent means ± SD; n = 3. F Western blotting of NSUN2 and SMO expression in SCs transfected with shCTL, shNSUN2, Vector, NSUN2-WT or NUSN2-MUT adenovirus. ACTB was used as a loading control. The relative protein expression was quantified by densitometry and normalized to ACTB. All data are shown as means ± S.D. G m⁵C-RIP-qPCR analysis of SMO in SCs transfected with shCTL, shNSUN2, Vector or NSUN2-WT adenovirus. Error bars, means ± S.D., n = 3. H Western blotting in control or SMO knockdown SCs transfected with NSUN2-WT or NSUN2-MUT adenovirus. ACTB was used as a protein loading control. The relative protein expression was quantified by densitometry and normalized to ACTB. All data are shown as means ± S.D. I Phase image in control or SMO knockdown SCs transfected with NSUN2-WT or NSUN2-MUT adenovirus. Scale bar: 100 μm. The P values were determined using Student’s t tests. *P < 0.05 **P < 0.01, ***P < 0.001