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. 2023 Jun 29;25(7):989–1003. doi: 10.1038/s41556-023-01163-3

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 5 — (a) Experimental plan and (b-c) representative IB for indicated proteins in (b) livers and (c) eWAT of 4–9 mo-old male mice injected with siCon, siMff or siDnm1l and fasted for 14–16 h (n = 4 mice). Quantifications for loss of MFF and DRP1 protein levels in liver are shown. (d) Frequency histograms depicting distribution of mitochondrial area and perimeter in Con, Rictor^KO and siMff or siDnm1l livers. N values for number of mice are in parenthesis: Con (n = 6), Rictor^KO (n = 6), siMff (n = 4), and siDnm1l (n = 4). Mean ± SEM is shown. (e) IB and quantification for indicated ER-stress and proteostasis markers in Hom and ER fractions from livers of 6 mo-old Con and Rictor^KO mice fasted for 14–16 h (n = 5 mice). (f-h) Representative IB for RICTOR in siCon or siRictor (f) AML12 or (g) HepG2 cells, or in (h) Rictor ^+/+ and Rictor ^-/- MEFs. RICTOR blots are representative of independent experiments obtaining similar results in AML12 (n = 4), HepG2 (n = 3), and MEF (n = 7) cells. (i) Representative confocal images of Rictor ^+/+ and Rictor ^-/- MEFs cultured in serum-free medium for 30 min in presence of MitoTracker green. Magnified insets are shown. Quantification for mitochondrial number and mitochondria size/shape descriptors is shown (Rictor^+/+ 36 cells and Rictor^-/- 43 cells from n = 3 independent experiments). (j) Representative IB for indicated proteins in Rictor^+/+ and Rictor^-/- MEFs (n = 3 independent experiments). Ponceau is loading control. Individual replicates and mean values are shown. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 are versus Con; ^#P < 0.05 versus Rictor^KO, two-tailed unpaired student’s t-test (b, e and i); One-way ANOVA followed by Tukey’s multiple comparisons test (d). Please refer to Supplementary Table 10_statistical summary. Source numerical data are available in SourceData_Table 1, and unprocessed blots are available in Source Data Extended data Fig. 5.

Source data