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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. 10 — (a) Significantly enriched interacting partners of CDC42 that belong to the RHO family of GTPases. A log2 transformed P-value cut-off of 4.32 was used as threshold (n = 4 independent experiments). (b) qPCR in NIH3T3 cells to validate the silencing of selected CDC42-binding partners identified by proteomics (n = 3 independent experiments). (c, d) IB and quantification for indicated proteins in Hom, Mp, MAMs, Cyt, and ER fractions from livers of (c) 5–6 mo-old Con or Rictor^KO male mice, and (d) 3–4 mo-old NDRG1^WT or NDRG1^Ser336Ala male mice co-injected with siRNA against endogenous Ndrg1 and fasted for 14–16 h. N values for number of mice analyzed for individual proteins in indicated fractions are in parentheses. For c, ARHGAP35: all fractions from Con (n = 4) and Rictor^KO (n = 4) mice; CDC42EP1: all fractions, Con (n = 6) and Rictor^KO (n = 8) mice except Rictor^KO Hom where n = 7 mice; ARHGDIA: all fractions from Con (n = 6) mice except Mp, MAMs and ER where n = 5 mice, and all fraction from Rictor^KO (n = 8) mice except Hom and ER where n = 7 mice. For d, ARHGAP35: all fractions from NDRG1^WT and NDRG1^Ser336Ala mice are n = 6 except Mp, MAMs and ER from NDRG1^WT mice where n = 5; CDC42EP1: all fractions from NDRG1^WT and NDRG1^Ser336Ala mice are n = 6 except MAMs in both groups where n = 5; ARHGDIA: all fractions from NDRG1^WT and NDRG1^Ser336Ala mice are n = 6 except Cyt in NDRG1^Ser336Ala mice where n = 5. Ponceau is loading control. Individual replicates and means are shown. *P < 0.05, ***P < 0.001 and ****P < 0.0001, two-tailed unpaired Student’s t-test (a-d). GAP: GTPase activating protein; GDI: GDP dissociation inhibitors. Please refer to Supplementary Table 10_statistical summary, and Supplementary Table 9. Source numerical data are available in SourceData_Table 1, and unprocessed blots are available in Source Data Extended Data Fig. 10.

Source data