Extended Data Fig. 5: Excessive overexpression of HIF1α is not able to rescue defective GSIS in Klb-null β-cells.
From: β-Klotho promotes glycolysis and glucose-stimulated insulin secretion via GP130
Islets isolated from 16-week-old male KlbRIP or KlbF/F mice were infected with adenovirus encoding Myc-tagged HIF1α (Adv-HIF1α) or GFP (Adv-GFP) at 300 multiplicity of infection (MOI) for 24 h. a, Representative immunoblots with anti-HIF1α, Myc, GFP or Tubulin antibodies in isolated islets infected with different adenoviruses (left) and quantification of HIF1α protein levels normalized to Tubulin (right, n = 6 mice). b-e, Insulin secretion (b, n = 8 mice), glycolytic flux measured by 3H2O generated from [5-3H]-glucose (c, n = 8 mice), mitochondrial TCA flux measured by 14CO2 generated from [2-14C]-pyruvate (d, n = 8 mice for KlbF/F-GFP, n = 8 mice for KlbRIP-GFP group, n = 9 mice for KlbRIP-HIF1α group) and ATP production (e, n = 6 mice) under basal (2.8 mM glucose) or high-glucose (16.7 mM) conditions in isolated islets infected with different adenoviruses. Data are presented as mean ± SEM. P values are derived from Welch’s ANOVA followed by Dunnett’s T3 multiple comparisons test (a), or ordinary two-way ANOVA followed by Tukey’s multiple-comparisons test (b-e).
