Fig. 5: PPARγ activation recovers adipocyte dysfunction caused by suppression of Rubicon.
From: Age-dependent loss of adipose Rubicon promotes metabolic disorders via excess autophagy
a Immunoblotting to detect adiponectin in Luciferase- or Rubicon-knockdown 3T3-L1 cells treated with or without 10 μM pioglitazone. Knockdown, with or without TZD treatment, was performed for 48 h starting on day 8 after induction of differentiation. n = 3 independent experiments. Quantification of adiponectin levels is shown in the graphs at right. b Relative mRNA expression of adipogenic genes in Luciferase- or Rubicon-knockdown 3T3-L1 cells treated with or without 10 μM pioglitazone. Knockdown, with or without TZD treatment, was performed for 48 h starting on day 8 after induction of differentiation. n = 3 independent experiments. c Relative triglyceride content in Luciferase- or Rubicon-knockdown 3T3-L1 cells treated with or without 10 μM pioglitazone. Knockdown, with or without TZD treatment, was performed for 72 h starting on day 8 after induction of differentiation. n = 3 independent experiments. d Representative images of ORO staining in Luciferase- or Rubicon-knockdown 3T3-L1 cells treated with or without 10 μM pioglitazone. Knockdown, with or without TZD treatment, was performed for 72 h starting on day 8 after induction of differentiation. Scale bars, 50 μm. n = 5 independent experiments. Quantification of relative ORO-stained area is shown in the graphs at right. Plasma adiponectin (e) and leptin (f) levels in control or Rubiconad−/− mice treated with or without 0.02% pioglitazone for 3 weeks. n = 7 mice (without TZD); n = 9 mice (with TZD). g Organ weight of eWAT, mWAT, iWAT, iBAT and liver from control or Rubiconad−/− mice treated with 0.02% pioglitazone for 3 weeks. n = 14 mice. Error bars indicate means ± SEM. Data were analysed by two-tailed Student’s t test (a–g). P value from left to right: 0.0009, 0.0650 (a), 0.0586, 0.3129, 0.5902, 0.0229, 0.2209, 0.7297, 0.9763 (b), 0.0002, 0.7647 (c), < 0.0001, 0.9078 (d), 0.0450, 0.8353 (e), 0.0441, 0.3403 (f), 0.8532, 0.2697, 0.8582, 0.0625, 0.9957 (g). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. N.S. not significant.
