Fig. 6. Mitochondrial proteomics identifies prohibitin 2 as a binding partner of COMP.

a Coomassie brilliant blue-stained gel showing bound proteins resolved by SDS-PAGE. The arrow indicates the bands that were excised and sequenced by mass spectrometry. b Co-immunoprecipitation assay of COMP and prohibitin 2 in rat VSMCs. The lysates were immunoprecipitated with anti-prohibitin 2 antibody, and the precipitates were analyzed by immunoblotting with COMP antibody. c Co-immunoprecipitation assay of COMP and prohibitin 2 in rat VSMCs. The lysates were immunoprecipitated with COMP antibody, and the precipitates were analyzed by immunoblotting with anti-prohibitin 2 antibody. d COS-7 cells were co-transfected with the pcDNA3.1-prohibitin 2 plasmid and flag-COMP plasmid, flag-COMP N-terminal plasmid, flag-COMP EGF domain plasmid, flag-COMP type-3 plasmid, or flag-COMP C-terminal plasmid. Proteins from cells were immunoprecipitated with flag antibody, and the precipitates were analyzed by immunoblotting with anti-prohibitin 2 antibody. e HEK293A cells were co-transfected with the Flag-COMP plasmid and 6 × His-prohibitin 2 N-terminal plasmid, 6 × His-prohibitin 2 PHB domain plasmid, or 6 × His-prohibitin 2 C-terminal plasmid. Proteins from cells were immunoprecipitated with the Flag antibody, and the precipitates were analyzed by immunoblotting with the His antibody