Adipose tissue dysfunction induces increased liver HIF-1α expression

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Fig. 5 Adipose tissue dysfunction induces increased liver HIF-1α expression. Adipose tissue dysfunction induces increased liver HIF-1α expression. (A) Liver interstitial oxygen tension (n = 4 mice per group). *P < 0.05, ***P < 0.001 versus lane 1. (B) HIF-α expression in primary hepatocytes incubated at different oxygen conditions for 3 hours. Arrowhead indicates nonspecific bands. (C) HIF-α expression in primary hepatocytes incubated with different combinations of leptin (100 ng/ml), insulin (100 nM), palmitic acid (400 μM), and fructose (5 mM) at three different concentrations of glucose (5, 10, and 21 mM) for 4 hours. (D) Dpp4 mRNA expression in WT and HIF-1α KO (HHKO) hepatocytes treated with leptin (100 ng/ml) + palmitic acid (PA) (400 μM) for 24 hours. *P < 0.05. (E) HIF-α expression in the liver of HFD WT mice treated with control immunoglobulin G (Ctl-IgG) or anti-leptin antibodies for 1 hour (n = 3 and 5 mice, respectively). (F) Western blot analysis of HIF-α and phosphorylated and total mTOR levels in primary hepatocytes incubated with different concentrations of leptin and/or adiponectin (AdipoQ) in the presence or absence of rapamycin (Rapa; 1 μM) for 3 hours. (G) Dpp4 mRNA expression in primary hepatocytes incubated with leptin (Lep) ± adiponectin for 16 hours. **P < 0.01 versus lane 1, #P < 0.01 versus lane 2. (H and I) HIF-1α protein expression (n = 5, 2, 2, and 3) (H) and Dpp4 mRNA levels (n = 3 samples per group) (I) in primary hepatocytes incubated with ACM of NCD or HFD WT mice for 3 hours (H) or 18 hours (I). For leptin neutralization, HFD ACM was incubated with control IgG or anti-leptin antibodies for 1 hour at 4°C before incubation with hepatocytes. **P < 0.05 versus lane 1, #P < 0.01 versus lane 2, $P < 0.05 versus lane 3. (J) Plasma leptin and adiponectin levels (n = 2, 9, and 9 mice per group). ***P < 0.001 versus lane 1, ###P < 0.001 versus lane 2. (K) Liver HIF-α expression. (L) Liver Hif1a and Glut1 mRNA expression (n = 10 mice per group). *P < 0.05. (M) Interstitial oxygen tension in the liver of HFD mice (n = 4 per group). (N) mRNA expression of Dpp4 in the liver of HFD mice (n = 10 mice per group). *P < 0.05. (O) Plasma intact/active GLP-1 levels in 6-hour–fasted HFD mice before or 10 min after glucose oral gavage (n = 7 mice per group). *P < 0.05. For statistical analysis, one-way (A, D, and G to J) or two-way (O) ANOVA with post hoc t tests between the individual groups or two-tailed unpaired t test (L and N) was performed. All data are presented as means ± SEM. Yun Sok Lee et al. Sci Adv 2019;5:eaaw4176 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).