Fig. 5: SIRT1 increased MTPα protein levels by inhibiting MTPα ubiquitylation. | Cell Death & Disease

Fig. 5: SIRT1 increased MTPα protein levels by inhibiting MTPα ubiquitylation.

From: SIRT1 deacetylates mitochondrial trifunctional enzyme α subunit to inhibit ubiquitylation and decrease insulin resistance

Fig. 5

a SIRT1 protein levels in 3T3-L1 adipocytes transfected with control siRNA, ovMTPα, or shMTPα. b MTPα protein levels in 3T3-L1 adipocytes transfected with control siRNA or shSIRT1. *p < 0.05 vs. shCON (t-test). c MTPα protein levels in 3T3-L1 adipocytes treated with EX-527 (10 μM) or resveratrol (10 μM) for 4 days. **p < 0.01 vs. control (one-way ANOVA). d MTPα mRNA levels in 3T3-L1 adipocytes transfected with control siRNA or shSIRT1. e MTPα mRNA levels in 3T3-L1 adipocytes treated with EX-527 (10 μM) or resveratrol (10 μM) for 4 days. f 3T3-L1 adipocytes treated with EX-527 (10 μM) or resveratrol (10 μM) for 4 days, then treated with CHX for the indicated time and subjected to western blotting and quantified. ***p < 0.001 vs. control (CHX 0); ####p < 0.001 vs. EX-527 (CHX 0) (two-way ANOVA). g 3T3-L1 adipocytes transfected with control siRNA or shSIRT1, treated with CHX for the indicated time and subjected to western blotting and quantified. **p < 0.01, ***p < 0.001 vs. control (CHX 0); #p < 0.001 vs. shSIRT1 (CHX 0) (two-way ANOVA). h MTPα protein levels in 3T3-L1 adipocytes treated with (+) or without (−) MG132 (10 μM) for 4 h. *p < 0.05 vs. MG132 (−) (t-test). i 3T3-L1 adipocytes treated with EX-527 (10 μM) or resveratrol (10 μM) for 4 days, or transfected with control siRNA or shSIRT1. Then MTPα was immunoprecipitated with anti-Ub antibody from 3T3-L1 adipocytes and ubiquitylation levels assessed by western blotting. **p < 0.01 vs. control (one-way ANOVA). ***p < 0.001 vs. shCON (t-test). j MTPα ubiquitylation and protein expression of SIRT1 and MTPα were measured in 3T3-L1 adipocytes transfected with control siRNA or shSIRT1 and treated with 9-PAHSA (20 μM) for 4 days. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control, #p < 0.05, ##p < 0.01 vs. 9-PAHSA (one-way ANOVA). k MTPα ubiquitylation measured in WAT of db/db mice after 10 days 9-PAHSA administration (50 mg/kg). **p < 0.01 vs. control (t test). Data represent at least three different experiments. n = 5 mice per group. All data represent means ± standard error (SE).

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