PI4KIIIα, EFR3B, and TTC7B form a complex at the plasma membrane. (A) Western blot analysis of anti-EFR3B and control immunoprecipitates from mouse brain homogenates. (B) Western blot analysis of anti-GFP, anti-HA, and anti-FLAG immunoprecipitates from lysates of HeLa cells transfected with GFP-M1-PI4KIIIα, EFR3B-HA, and TTC7B-FLAG. (C) Confocal imaging of live HeLa cells transfected with TTC7B-GFP (top) or EFR3B-tdTomato (bottom). (D) Imaging analysis of HeLa cells triple transfected with GFP-M1-PI4KIIIα, TTC7B-mCherry, and EFR3B-HA. EFR3B-HA immunofluorescence was performed with an anti-HA antibody followed by an Alexa Fluor 647–conjugated anti–rat secondary antibody. (E and F) Imaging analysis of HeLa cells double transfected with EFR3B-HA and either TTC7B-mCherry (E) or GFP-M1-PI4KIIIα (F) after anti-HA immunofluorescence labeling. Insets show the region outlined with dotted lines at higher magnification. Bars: (full size images) 20 µm; (insets) 2 µm.