Extended Data Fig. 1: WAT-selective upregulation of mitochondrial protein turnover and mitochondrial proteostasis regulators.
From: Proteolytic rewiring of mitochondria by LONP1 directs cell identity switching of adipocytes
(a, b) 5-week-old WT mice exposed to 8˚C or 30˚C for 7 days. (a) Body temperature and iWAT mass measurements. (b) Representative UCP1 staining and H&E staining of iWAT section after exposure to cold or thermoneutrality. Data shown are representative of 6 (a) or 5 (b) mice/group. (c) iWAT and BAT MitoTimer Red:Green ratio of Adipoq-Cre/MitoTimer mice exposed to 8°C or 30°C. > 38 images counted from 7 mice/group. (d, e) 4-week-old WT mice were injected with CL316,243 for 5 days. (d) Oxygen consumption measurements before and after injection. n = 8 mice/genotype. (e) Representative UCP1 staining and H&E staining of iWAT section. n = 8 mice/genotype. Scale bar, 50 μm. (f) iWAT MitoTimer Red:Green ratio of MitoTimer mice at indicated time points of CL316,243-treatment. >48 images counted from 5 mice/group. (g) BAT MitoTimer fluorescence images at indicated time points of CL316,243-treatment and quantification (h). Scale bar, 10 μm. Representative images are shown. >30 images counted from 5 mice/group. (i) LONP1/Tubulin and CLPP/Tubulin protein ratios in iWAT and BAT of WT mice exposed to 8°C or 30°C. n = 8 mice/group. (j, k) Immunoblot analysis of proteins in EWAT isolated from mice after thermogenic stimulations. n = 4 mice/group. (l) Immunoblot analysis of proteins in iWAT of WT mice exposed to 8°C or 30°C for 7 days. n = 4 mice/group. (m) iWAT Mitochondrial DNA levels of WT mice exposed to cold or thermoneutrality. n30˚C = 10; n8˚C = 12 mice. (n) Thermogenic gene expression levels in differentiated beige adipocytes. n = 5 experiments. (o) MitoTimer Red:Green ratio in adipocytes transfected with Lonp1 or Clpp siRNA. >77 images counted from 5 experiments. (p) MitoTimer Red:Green ratio in WT and LONP1 AKO iWAT adipocytes differentiated with or without Rosi. >59 images counted from 6 experiments. All data are shown as mean ± SEM. P value was calculated by unpaired two-sided Student’s t-test (a, c, i-o) or one-way ANOVA (f, h) or two-way ANOVA (d, p). Source numerical data and unprocessed blots are available in source data.
