Fig. 6: Psmb8 strongly associated with Drp1.

A The MS/MS spectrum and extracted ion chromatograms from LC-MS/MS analysis of the derived Drp1 peptide. B Column chart showing the statistical proteins of LC-MS/MS analysis results. C The association between full-length Psmb8 and full-length Drp1 proteins was predicted by the ZDOCK server. Their structures were visualised using PyMOL software. D The association of endogenous Psmb8 with Drp1 protein in heart lysates, which were immunoprecipitated (IP) with IgG control or anti-Psmb8 antibody and then detected by immunoblotting analysis with anti-Drp1 or anti-Psmb8 antibody, respectively. E Analysis of the interaction between endogenous Psmb8 and Drp1 proteins in NRCM lysates treated with/without MG132 (20 μM) by IP with IgG control or anti-Drp1 antibody following by immunoblotting with antibody against Drp1 or Psmb8 under/not under H/R, respectively. F Fluorescence localisation of the Psmb8 and Drp1 proteins in NRCMs. G Identification of the Drp1-binding domains within the Psmb8. Whole cell lysates isolated from HEK293 cells co-transfected with Flag-tagged full-length Psmb8 or Flag-GFP-tagged Psmb8 mutants and Myc-tagged Drp1 plasmids were IP with anti-Myc antibody and analysed by immunoblotting with anti-Flag or Myc antibody. H Mapping of the Psmb8-binding domains within the Drp1. Whole cell lysates isolated from HEK293 cells co-transfected with Flag-tagged full-length Psmb8 and Myc-tagged full-length Drp1 or mutant plasmids were IP with anti-Flag antibody followed by immunoblotting with anti-Myc or Flag antibody. I Schematic diagram for the amino acids (69-276) of the Psmb8 binding to Drp1. J Schematic diagram for the amino acids (302-501) of Drp1 binding to Psmb8. K Prediction of the interaction between Psmb8 C-terminal fragment (73-275 aa) and the truncated Drp1 proteins (1-301aa, and 302-736 aa) by the ZDOCK server. The predicted structure was visualised by PyMOL software.