Fig. 2

FAM177A1 is a protein neighbor of VPS13B in the Golgi complex. (A) Alphafold2 prediction of human FAM177A1 structure. (B) COS7 cells expressing VPS13B^Halo, FAM177A1-GFP, and GalT-RFP showing that FAM177A1 is localized at the Golgi complex. (C) Western blot analysis of soluble cytosolic fraction and total membrane faction of HeLa cells expressing FAM177A1-GFP. GAPDH was used as a soluble control, Stx6 as a membrane control protein. (D) Top panel: Co-expression of FAM177A1-helix1-GFP and GalT-RFP in COS7 cells. Bottom panel: Co-expression of FAM177A1-?helix 1-GFP and GalT-RFP in COS7 cells. Scale bar = 10 µm. (E) FLASH-PAINT performed in HeLa cells expressing VPS13B^GFP and FAM177A1-Halo and immunolabeled with anti-GFP, anti-halo, and anti-GM130 antibodies. (F) Median distances between super-resolved signals of different Golgi complex targets. Only signals closer than 500 nm to each other were considered. (G) Snapshots of COS7 cells expressing FAM177A1-SNAP, ZFPL1-GFP, and GalT-RFP treated with water for 10 min. Scale bar = 5 µm. (H) Snapshots of HeLa cells expressing FAM177A1 GFP and VPS13B^Halo before and after BFA (5 µg/ml for 50 min) treatment. Scale bar = 10 µm. Source data are available for this figure: SourceData F2.