Extended Data Fig. 6: SDH-mediated succinate metabolism is critical for LONP1-regulated cell fate programming of mature adipocytes.
From: Proteolytic rewiring of mitochondria by LONP1 directs cell identity switching of adipocytes
(a) UCP1 staining of BAT sections from WT mice treated with vehicle or CL, fed with or without 1.5% sodium succinate for 2 weeks. Scale bar, 100 μm. Images represent 4 mice/group. (b) UCP1/Tubulin signal ratios normalized to WT vehicle-treated mice as described in Fig. 6b. nCon = 6; nCL = 6; nCon+Succinate = 6; nCL+Succinate = 7 mice (c) Schematic illustration of AAV-mediated SDHB overexpression experiment. UCP1 and H&E staining of iWAT sections after treatments. Representative images are shown. nGFP Con = 10; nGFP CL+succinate = 9; nSDHB Con = 9; nSDHB CL+Succinate = 8 mice. Scale bar, 50 μm. (d) Immunoblot analysis of proteins in iWAT. n = 8 mice/group. (e) iWAT mitochondrial respiration rates. nGFP Con = 9; nGFP CL+succinate = 10; nSDHB Con = 9; nSDHB CL+Succinate = 10 mice. (f) Malonate inhibition of SDH experiment schematic. UCP1 and H&E staining of iWAT sections. Representative images are shown. nCon PBS = 10; nCon CL = 11; nMalo PBS = 5; nMalo CL = 7 mice. Scale bar, 50 μm. (g-i) Immunoblot analysis (g) (nCon PBS = 10; nCon CL = 10; nMalo PBS = 9; nMalo CL = 10 mice), mitochondrial respiration rates (h) (nCon PBS = 8; nCon CL = 9; nMalo PBS = 8; nMalo CL = 9 mice) and succinate content (i) (nCon PBS = 7; nCon CL = 10; nMalo PBS = 7; nMalo CL = 10 mice) in iWAT from WT mice of indicated treatments. (j) AdipoChaser mice treated with doxycycline-diet for 10 days, followed by CDDO alone and in combination with malonate-mediated SDH inhibition for 12 days. (k) Representative image of iWAT sections stained with anti-GFP (green) and anti-PERILIPIN (red) antibodies and counterstained with DAPI (blue). nPluse = 4; nControl = 6; nCL = 4; nCDDO+CL = 5; nCDDO+CL+Succinate = 6 mice. Scale bar, 20 μm. (l) Percentage of multilocular adipocytes from labeled cells (expressing mGFP), nPluse = 4; nControl = 6; nCL = 4; nCDDO+CL = 5; nCDDO+CL+Succinate = 6 mice. >400 adipocytes were counted for each condition. All data are shown as mean ± SEM. P value was calculated by two-way ANOVA (b, d, e, g-i) or one-way ANOVA (l). Source numerical data and unprocessed blots are available in source data.
