Extended Data Fig. 4: Evaluation of the effects by LONP1 inactivation on creatine- or SERCA-mediated thermogenic pathways.
From: Proteolytic rewiring of mitochondria by LONP1 directs cell identity switching of adipocytes
(a) Immunoblot analysis on the indicated proteins in BAT from 5-week-age WT and Lonp1f/f/UCP1-Cre mice. n = 6 mice per group. (b) H&E staining of BAT sections from 5-week-old WT and Lonp1f/f/UCP1-Cre mice. Images are representative of 5 mice per group. (c, d) Representative UCP1 and H&E staining of iWAT sections from 5-week-old WT and Lonp1f/f/UCP1-Cre mice subjected to cold exposure (c: nWT 30˚C = 4; nWT 8˚C = 5; nUCP1-Cre 30˚C = 5; nUCP1-Cre 8˚C = 4 mice) or β3-AR agonist CL316,243 treatment (d: nWT Con = 5; nWT CL = 5; nUCP1-Cre Con = 4; nUCP1-Cre CL = 5 mice). Scale bar, 50 μm. (e) Body temperature of 5-week-old WT and Lonp1f/f/UCP1-Cre mice exposed to cold (4˚C) for 6 hours. nWT = 5; nUCP1-Cre = 6 mice per group. (f) RT-qPCR analysis of expression of the genes involved in creatine- and SERCA-mediated thermogenesis in WT and LONP1 AKO adipocytes, n = 5 independent experiments. (g) Immunoblot analysis of CKB, GAMT, CKMT1A and LONP1 protein levels (left) and quantification of CKB/Tubulin, GAMT/Tubulin and CKMT1A/Tubulin protein ratios (right) in WT and LONP1 AKO adipocytes. n = 6 independent experiments. (h) Creatine and PCr to creatine (Cr) ratio in WT and LONP1 AKO adipocytes. n = 5 independent experiments. (i, j) RT-qPCR analysis of expression of the genes involved in creatine- and SERCA-mediated thermogenesis in iWAT (i: nWT 30˚C = 7; nWT 8˚C = 7; nAKO 30˚C = 5; nAKO 8˚C = 7 mice) and BAT (j: nWT 30˚C = 8; nAKO 30˚C = 9; nWT 8˚C = 8; nAKO 8˚C = 8 mice) of WT and LONP1 AKO mice exposed to cold or thermoneutrality. All data are shown as the mean ± SEM. P value was calculated by unpaired two-sided Student’s t-test (f, g, h) or two-way ANOVA (e, i, j). Source numerical data and unprocessed blots are available in source data.
