Fig. 4: FTOrs9939609-A causes an increased propensity for HFD serum-induced insulin resistance. | Nature Communications

Fig. 4: FTOrs9939609-A causes an increased propensity for HFD serum-induced insulin resistance.

From: An obesogenic FTO allele causes accelerated development, growth and insulin resistance in human skeletal muscle cells

Fig. 4

a A schematic diagram illustrating the workflow by which HFD serum was tested for the induction of insulin resistance in the human myocytes model. b Left: representative images of FoxO1-GFP-infected FTOrs9939609-TT-myocytes and FTOrs9939609-A-myocytes exposed to 1% HFD serum at day 0 and day 14. Scale bars, 200 μm. Middle: quantification of percentage of FoxO1-GFP+ nuclei numbers at the end of day 14, n = 3 biologically independent samples. Right: quantification of nuclear/cytoplasmic FoxO1-GFP ratios at the end of day 14, n = 3 biologically independent samples. c Left: western blot of phospho-IRS1, phospho-AKT, phospho-FOXO and GAPDH protein abundance in FTOrs9939609-TT-myotubes: H1, H7, H9 and FTOrs9939609-A-myotubes: FTO-H1-1, FTO-H1-2, FTO-H7-1, FTO-H7-2, FTO-H9-1, FTO-H9-2 exposed to 1% HFD serum at day 14. Right: quantification of the abundance of insulin signaling proteins, n = 3 biologically independent samples. d Measurement of glucose uptake in FTOrs9939609-TT-myocytes and FTOrs9939609-A-myocytes in the presence of 1 µg/ml insulin or not (FTO-TT + 2-DG(-)/Insulin(-) vs. FTO-A + 2-DG(-)/Insulin(-), P = 0.72198; FTO-TT + 2-DG(+)/Insulin(-) vs. FTO-A + 2-DG(+)/Insulin(-), P = 0.23893; FTO-TT + 2-DG(+)/Insulin(+) vs. FTO-A + 2-DG(+)/Insulin(+), P = 0.03833), n = 3 biologically independent samples. e Left: Western blot of phospho-IRS1, phospho-AKT, phospho-FOXO and GAPDH protein abundance in FTOrs9939609-TT-myotubes: H1, H7, H9 and FTOrs9939609-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 insulin signaling proteins, n = 3 biologically independent samples. f Left: Western blot of phospho-AKT, phospho-FOXO and GAPDH protein abundance in FTOrs9939609-TT-myotubes: H1, H7, H9 and FTOrs9939609-A-myotubes: FTO-H1-1, FTO-H1-2, FTO-H7-1, FTO-H7-2, FTO-H9-1, FTO-H9-2, under serum-fed (Fed), 18 hr serum starved (Ss) or insulin-stimulated (Ins) conditions. Insulin stimulation was performed in serum-starved myotubes with 10 μg/mL insulin for 5 min. Right: quantification of the abundance of insulin signaling 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 (a). 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.

Back to article page