Fig. 1: Amongst different lineages, FTO^rs9939609-A promotes myogenesis most robustly.

a A schematic diagram illustrating our workflow utilizing CRISPR/Cas9-based prime editing of hESCs to study the effects of a specific FTO SNP on cellular differentiation. b Quantitative RT-PCR of myogenic markers in FTO^rs9939609-TT and FTO^rs9939609-A hESCs-derived myogenic progenitors, myocytes and myotubes. Paired box 3 (PAX3), Paired box 7 (PAX7), Myogenic factor 5 (MYF5), Myogenic differentiation 1 (MYOD1), Myogenin (MYOG), Skeletal muscle actin alpha 1 (ACTA1), Myosin heavy chain (MYHC), Neural cell adhesion molecule 1 (NCAM1), Actin Beta (ACTB), n = 3 biologically independent samples. c Gene Set Enrichment Analysis (GSEA) of the signatures enriched in FTO^rs9939609-A-myotubes, relative to FTO^rs9939609-TT -myotubes, according to RNA-sequencing. d Left: western blot quantification of FTO, PAX3, PAX7, MYOD1, MYOG, MHC (myosin heavy chain, MF20) 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 myogenic differentiation proteins, 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. Data were analyzed using Wald test (c). 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.