Fig. 3: CLASP confidently localizes FAM136A to the IMS.

A Cross-link map of FAM136A and its interacting proteins. LMs are shown in green; FAM136A is shown in purple. The CLASP annotation of FAM136A is supported by direct connections to 1 IMS LM and 3 IMM LMs. Proteins that are not LMs but support the IMS localization of FAM136A are shown in grey. “Predict region” indicates the protein/protein region, for which a CLASP prediction was made. B Confocal fluorescence images of C-terminally HA-tagged FAM136A (FAM136A-HA) and OMM marker TOMM20 in HeLa cells (left panel). Images of untransfected cells are shown as a negative control in the bottom left. The color bar shows the contrast setting. Scale bars = 10 μm. C STED microscopy images of FAM136A-HA-transfected HeLa-COX8A-SNAP cells stained with BG-SiR-d12 (to label the IMM marker COX8A) and HA antibody. Scale bar = 1 μm. D. Alkaline carbonate extraction of mitochondria isolated from HEK293T cells overexpressing FAM136A-HA. The OMM protein TOMM70, IMM protein TIMM23 and IMM associated protein TIMM44 are used as markers for each sub-compartment. T, total mitochondrial extraction; S, supernatant; P, pellet of mitochondrial membrane. E Protease protection assay to analyze the localization of FAM136A-HA in HEK293T cells. OMM protein TOMM70, IMM protein TIMM23 and matrix protein MRPS18B are used as markers for each mitochondrial sub-compartment. Experiments in panels B-E were performed once. Source data are provided as a Source Data file.