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. Author manuscript; available in PMC: 2022 Mar 2.
Published in final edited form as: Cell Metab. 2021 Feb 4;33(3):531–546.e9. doi: 10.1016/j.cmet.2021.01.005

Figure 5. C9orf72 Recruits the PHB Complex to Stabilize TIMMDC1.

Figure 5.

(A) Co-IP of PHB1 and PHB2 with anti-C9orf72 antibody was analyzed with isolated mitochondria from WT and C9KO MEFs

(B) Mitochondria isolated from HEK293 cells were solubilized and subjected to immunoprecipitation with the anti-C9orf72 antibody, followed by immunodetection of PHB2.

(C) Mitochondria lysates from HEK293 cells were subjected to immunoprecipitation with the anti-PHB2 antibody, followed by immunodetection of C9orf72.

(D) The direct interaction of purified His-SUMO-C9orf72 and GST-PHB2 was determined by the Ni-NTA pull-down of C9orf72, followed by immunoblot analysis of the proteins.

(E) WT and C9KO MEFs were transduced with lentiviral PHB2 shRNAs and the knockdown efficiency of PHBs was determined by immunoblotting. CTRL: non-targeting control shRNA.

(F) CHX chase analysis of the turnover of TIMMDC1 in WT and C9KO MEFs, with or without the knockdown of PHB2. Quantifications of the turnover rates of TIMMDC1 are shown (n = 4).

Data are means ± s.d., analyzed by one-way analysis of variance (ANOVA). ** P < 0.01; *** P < 0.001; ns, not significant. See also Figures S7 and Table S5.