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. 2017 Jun 1;7:2661. doi: 10.1038/s41598-017-02838-2

Figure 6.

Figure 6

Increase in glucose metabolism induced by GLP-1R signalling is mediated by HIF-1α. (A) BRIN-BD11 cells were treated as indicated for 18 hours and Hif1α protein levels were determined by immunoblot analysis. (B) After treatment with Exendin-4 for 18 hours HIF-1α mRNA expression was evaluated by qRT-PCR. (C) BRIN-BD11 cells were pre-incubated with either vehicle, 1 µM Torin 1 or 50 µM LY294002 for 30 min as indicated, followed by addition (or not) of 50 nM Exendin-4 for 18 hours. Immunoblot shows Hif1α protein expression and phosphorylation levels of mTOR at serine 2448. (D–J) BRIN-BD11 cells were transfected with either HIF-1α specific siRNAs (siHIF1a) or non-targeting siRNAs (siControl) for 24 h, and subsequently exposed to 50 nM Exendin-4 (or not) for 18 h. (D) Exendin-4’s ability to promote protein levels of Pdh and Pfkp was significantly blunted in cells transfected with siHIF1a as compared to siControl transfected cells. (E) At the mRNA level, siHIF1a transfection caused a reduction of approximately 80% in HIF-1α expression. (F) mRNA expression of glycolytic enzymes was determined by qRT-PCR analysis. (G–J) Determination of glucose consumption, 2-NBDG uptake, SSC-A and FSC-A following HIF-1α knockdown. Data are mean ± SEM, n ≥ 3; n.s. = non-significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.001.