The CHCH domain is necessary for mitochondrial import of CHCHD10 The CHCH domain is necessary for mitochondrial import of CHCHD10 HeLa cells were transfected with the indicated CHCHD10 variants (D10‐HA) and GFP‐fusion proteins. A–D(A, D) Double immunofluorescence using ATP5A1 as a mitochondrial marker protein. Cells with similar expression level are shown. Scale bars represent 10 μm. (B, C) Representative immunoblot of biochemical fractionation of mitochondria and cytosol using antibodies against HA, ATP5A1, and actin followed by quantitative analysis of the respective CHCHD10 truncation mutant. Levels of HA‐tagged CHCHD10 (D10‐HA) were either normalized to ATP5A1 (for mitochondria) or actin (for cytosol). Data are shown as mean ± SD. One‐way ANOVA (followed by Dunnett's post hoc test against WT): n = 3–4 biological replicates, mitochondrial: WT versus ΔCHCH **P = 0.0013, WT versus Q108* **P = 0.0019, WT versus Q108P **P = 0.0030, WT versus ΔNT‐Q108P **P = 0.0020; Cytosolic: WT versus Q108* *P = 0.0428.Source data are available online for this figure. Carina Lehmer et al. EMBO Mol Med. 2018;10:e8558 © as stated in the article, figure or figure legend