Analysis of the interaction of ARV1 with CD55 or PIGQ. (A) Schematic model of the BiFC system. Split Citrines (N-fragment and C-fragment) were fused at C-termini or N-termini of ARV1, PIGQ, CD55(C), and PIGH. When the N-fragment and C-fragment of Citrine interact with each other, mature fluorescence protein is generated. (B) Fluorescence images of PIGS-HRD1-CD55-TKO cells transiently coexpressing RFP-KDEL and split Citrine fused proteins. (C) The mean value of the fluorescence intensity of cells shown in B was calculated by ImageJ and displayed as the mean (Yellow/Red) ± SD of five independent cells. (D) Cell lysates prepared from PIGS-HRD1-CD55-TKO transfectants were immunoprecipitated with anti-Flag beads. Samples were analyzed by WB. Proteins were detected using the Flag or Myc antibodies. (E) Cell lysates prepared from PIGS-HRD1-CD55-TKO transfectants were immunoprecipitated with anti-Flag beads. Samples were analyzed by WB. Proteins were detected using the Flag, HA, or Myc antibodies. Coimmunoprecipitated level was indicated with numbers in red. Source data are available for this figure: SourceData FS5.