Panel A: Western blot analysis showing interaction between T7-MUNC13-4 and FLAG-RAB27A. The blot includes four lanes labeled Mock, FLAG-RAB27A(WT), and FLAG-RAB27A(S115R). The top blot shows T7-MUNC13-4 after immunoprecipitation with anti-FLAG, the second blot shows T7-MUNC13-4 input, the third blot shows FLAG-RAB27A after immunoprecipitation with anti-FLAG, and the bottom blot shows FLAG-RAB27A input. Panel B: Western blot analysis showing interaction between T7-MLPH and FLAG-RAB27A. The blot includes four lanes labeled Mock and T7-MLPH with FLAG-RAB27A(WT) and FLAG-RAB27A(S115R). The top blot shows FLAG-RAB27A after immunoprecipitation with anti-T7, the second blot shows FLAG-RAB27A input, the third blot shows T7-MLPH after immunoprecipitation with anti-T7, and the bottom blot shows T7-MLPH input. Molecular mass markers in kilodaltons are shown on the left.
RAB27A effector-binding activities of RAB27A(Ser115Arg). (A) Interaction between T7-MUNC13-4 and FLAG-RAB27A(WT or Ser115Arg; indicated as S115R). (B) T7-MLPH and FLAG-RAB27A(WT or Ser115Arg). These protein interactions in COS-7 cell lysates were analyzed by co-immunoprecipitation assays using anti-FLAG tag antibody-conjugated agarose beads (A) or anti-T7 tag antibody-conjugated agarose beads (B). Co-immunoprecipitated T7-MUNC13-4 (or FLAG-RAB27A) and immunoprecipitated (IP) FLAG-RAB27A (or T7-MLPH) were detected by immunoblotting with HRP-conjugated anti-T7 tag antibody and anti-FLAG tag antibody (or anti-FLAG tag antibody and anti-T7 tag antibody), respectively. The positions of the molecular mass markers (in kilodaltons) are shown on the left. Data in A and B are representative of the data obtained in three independent experiments, and similar results were obtained in each experiment. Source data are available for this figure: SourceData F5.