Figure 1: BARS interacts with LPAATδ.
(a) Representative confocal microscopy images of COS7 cells transfected with Flag-tagged LPAATγ, LPAATδ and LPAATη, and fixed and processed for immunofluorescence with a monoclonal anti-Flag antibody (green) and with a polyclonal anti-TGN46 antibody (red; as indicated). Dotted lines indicate cell borders. (b) BARS immunoprecipitation (IP:BARS) of lysate from HeLa cells co-expressing BARS and LPAATγ–Flag, LPAATδ–Flag or LPAATη–Flag. Representative western blotting (antibodies as indicated) of total lysate (input) and immunoprecipitated proteins with preimmune-IgG (Preim-IgG) or anti-BARS-IgG (as indicated). IgGH, IgG heavy chain. (c) Immunoprecipitation with an anti-Flag antibody (IP:Flag) of lysate from HeLa cells co-transfected with BARS and LPAATγ–Flag, LPAATδ–Flag or the empty vector. Representative western blotting of total lysate (input) and Flag-immunoprecipitated proteins with an anti-Flag monoclonal antibody or the anti-BARS polyclonal antibody (as indicated). (d) Representative electron microscopy image of HeLa cells transfected with Flag-tagged LPAATδ for 24 h, and fixed and processed for cryo-immuno-electron microscopy with a monoclonal anti-Golgin-97 antibody (15-nm gold particles) and with a polyclonal anti-LPAATδ antibody (10-nm gold particles). Molecular weight standards (kDa) in b and c, are indicated on the left of each panel. Data are representative of three independent experiments. Scale bars, 10 μm (a); 200 nm (d).
