Extended Data Fig. 8: Mini-sized TC tagged VDAC3 ensures its localization on mitochondrial surface for imaging experiments.

a, 4 plasmid constructs of VDAC3. N- or C-termini were fused with TC tag or APEX2 protein. b, Mini-sized TC tag and its self-labeling mechanism by FlAsH-EDT₂ fluorescent reporter for imaging experiments. c, N-terminal fusion or C-terminal fusion of VDAC3 with mini-sized TC tag ensured its localization to mitochondrial surface (3#, 4# lanes). The large-sized APEX2 protein fusion with VDAC3 mis-localized VDAC3 to nucleus or formed aggregates in the cell (1#, 2# lanes), impeding its application for interaction assay. Green: FlAsH signal. Red: mitochondria stained by Mito-TrackerTM Deep Red. d, Co-localization of VDAC3 labeled by FlAsH and LDs stained by B6, indicating close contact between LDs and VDAC3. e, Co-localization of VDAC3 labeled by FlAsH and PLIN3 tagged by mCherry, indicating close proximity between PLIN3 and VDAC3. Images are representative of twice independent experiments (c-e). f, Schematic of split GFP complementation assay.