Fig. 5: GCC88 and golgin-97 share a distinct pool of TMEM87A and TGN46 enriched vesicles.

a PCA projections for the LOPIT-DC of mito versus GCC88-mito showing SVM classifications and the top six hits that are localised to the endosomal network from Fig. 1f. b TMT reporter ion distributions of proteins across fractions for each replicate for mito, golgin-97-mito and GCC88-mito showing the profiles of known mitochondrial markers (Mitochondria) and the indicated proteins. c Confocal micrographs of HeLa cells expressing BioID-mito and the indicated golgin-mito constructs (HA) stained for endogenous TMEM87A (cargo). Micrographs representative of three independent experiments, scale bars 10 μm. d Correlative light and electron tomography of resin-embedded HeLa cells co-transfected for 24 hours with golgin-97-mito and TMEM87A-RFP (green), and stained with mitotracker DeepRed (magenta) to label mitochondria. Left: Fluorescence micrograph of a section of a resin-embedded cell. White circles indicate TMEM87A-RFP signals localised in the electron tomogram. Middle-left: Virtual slice from the electron tomogram acquired at the area indicated with a white square on the left image—green crosses indicate predicted positions of the TMEM87A-RFP signal centroids shown on the fluorescence image. Tomogram representative of two independent replicates. Middle: magnified area from white square in the tomogram slice showing vesicles accumulated between two mitochondria, with two views of a 3D segmentation model of this same area (mitochondria in purple and vesicles in green). Full tomogram and rendering in Supplementary Movie 1.