Fig. 5: FTO^rs9939609-A induces an increase in IGF2 and H19 via m⁶A demethylation.

a Left: Volcano plot of differentially expressed genes in FTO^rs9939609-A-myotubes, relative to FTO^rs9939609-TT-myotubes, according to RNA-sequencing. Right: Gene Set Enrichment Analysis (GSEA) of the IGF signature enriched in FTO^rs9939609-A-myotubes, relative to FTO^rs9939609-TT-myotubes. b ELISA of IGF2 protein secretion by FTO^rs9939609-TT-myotubes and FTO^rs9939609-A-myotubes into the culture medium after exposure to 1% HFD serum for 1 day, 3 days, 7 days, and 14 days, n = 3 biologically independent samples. c Left: Western blot of IGF2 and GAPDH protein abundance in FTO^rs9939609-TT-myotubes: H1, H7, H9 and FTO^rs9939609-A-myotubes: FTO-H1-1, FTO-H1-2, FTO-H7-1, FTO-H7-2, FTO-H9-1, FTO-H9-2. Right: quantification of the abundance of IGF2 and GAPDH proteins (FTO-TT vs. FTO-A, P = 0.00099), n = 3 biologically independent samples. d Quantitative RT-PCR for FTO mRNA, IGF2 mRNA and H19 lncRNA gene expression in FTO^rs9939609-TT-myotubes and FTO^rs9939609A-myotubes, n = 3 biologically independent samples. e Encode database: cis-Regulatory Elements near rs9939609 in skeletal muscle cells and skeletal muscle tissues. f H3K27Ac ChIP-qPCR of EH38E1816455 enhancer in FTO^rs9939609-TT-myotubes and FTO^rs9939609-A-myotubes (FTO-TT vs. FTO-A, P = 0.00019), n = 3 biologically independent samples. g N6-methyladenosine (m⁶A) MeRIP-qPCR for IGF2 and H19 in FTO^rs9939609-TT-myotubes and FTO^rs9939609-A-myotubes, n = 3 biologically independent samples. h Quantitative RT-PCR for IGF2 and H19 lncRNA gene expression in FTO^rs9939609-TT-myotubes, treated with DMSO, 200 nM bisantrene or 50 μM entacapone, n = 3 biologically independent samples. H1 hESC line colored in purple, H7 hESC line colored in light blue and H9 hESC line colored in dark blue. Data are presented as mean + SEM. P values were calculated by two-tailed unpaired t-test. *P < 0.05, **P < 0.01. Source data are provided as a Source Data file.