Fig. 6: ERα-derived AF1 peptide increases hepatic insulin sensitivity through IRS1.
From: An estrogen receptor α-derived peptide improves glucose homeostasis during obesity
a SVM score of interaction between IRS1 and ERα domains. b Effect of ERα 1-60 domain and AF1 peptide (34 aa) on insulin sensitivity in HepG2 cells, n = 3 independent cells; for IRS1, Insulin versus 34 aa peptide-Insulin, P = 0.0046, Insulin versus ERα 1-60-Insulin, P = 0.0006; for pAKT-S473, Vehicle versus Insulin, P < 0.0001, Insulin versus 34 aa peptide-Insulin, P = 0.0044, Insulin versus ERα 1-60-Insulin, P = 0.0030. c Effect of ERα 60-280 domain on insulin sensitivity in HepG2 cells, n = 3 independent cells; for pAKT-S473, Vehicle versus Insulin, P < 0.0001, ERα 60-280-Vehicle versus ERα 60-280-Insulin, P < 0.0001. d Immunofluorescence staining of AF1 peptide (34 aa) and ERα 1-60 domain in HEK293T cells. The experiments were repeated independently twice. Representative images were shown. e Amino acid sequence of FITC labeled AF1 peptide conjugated with TAT. f Interaction between AF1 peptide and IRS1 protein domains. The experiments were repeated independently twice. Representative results were shown. g Effect of AF1 peptide on IRS1 phosphorylation at S302 in primary mouse hepatocytes, n = 3 independent cells; for pIRS1-S302, P = 0.0377; for IRS1, P = 0.0156. h Effect of AF1 peptide on IRS1 ubiquitination in HEK293T cells. The experiments were repeated independently twice. Representative blots were shown. i Effect of AF1 peptide on insulin-induced suppression of HGP in primary hepatocytes upon glucagon treatment, n = 4 independent cells; CNTR-Vehicle versus CNTR-Glucagon, P = 0.0006, AF1 peptide-Vehicle versus AF1 peptide-Glucagon, P = 0.0272, CNTR-Glucagon-Insulin versus AF1 peptide-Glucagon-Insulin, P = 0.0019. j Effect of AF1 peptide on insulin sensitivity in primary mouse hepatocytes, n = 3 independent cells; for IRS1, CNTR-Vehicle versus AF1 peptide-Vehicle, P = 0.0067, CNTR-Insulin versus AF1 peptide-Insulin, P = 0.0314, for pAKT-S473, AF1 peptide-Vehicle versus AF1 peptide-Insulin, P = 0.0001, CNTR-Insulin versus AF1 peptide-Insulin, P = 0.0008; for pAKT-T308, CNTR-Vehicle versus CNTR-Insulin, P = 0.0154, AF1 peptide-Vehicle versus AF1 peptide-Insulin, P = 0.0002, CNTR-Insulin versus AF1 peptide-Insulin, P = 0.0167. k Effect of AF1 peptide on insulin sensitivity in control and ERα deficient primary mouse hepatocytes, n = 3 independent cells, for IRS1, ERαF/F-Insulin versus ERαLivKO-Insulin, P < 0.0001, ERαLivKO-Insulin versus ERαLivKO-Insuin-AF1 peptide, P = 0.0061, for pAKT-S473, ERαF/F-Vehicle versus ERαF/F-Insulin, P < 0.0001, ERαF/F-Insulin versus ERαLivKO-Insulin, P < 0.0001, ERαLivKO-Insulin versus ERαLivKO-Insulin-AF1 peptide, P = 0.0034; for pAKT-T308, ERαF/F-Vehicle versus ERαF/F-Insulin, P < 0.0001, ERαF/F-Insulin versus ERαLivKO-Insulin, P < 0.0001, ERαLivKO-Insulin versus ERαLivKO-Insulin-AF1 peptide, P = 0.0090. Data are presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, unpaired Two-tailed Student’s t test (g), One-way ANOVA (b, i, k), Two-way ANOVA with Tukey’s multiple comparisons test (c, j). CNTR: Control. Source data are provided as a Source Data file.
