Fig. 3: TRIM31 interacted with P53.

a The diagram showed the process for the quantitative proteomics and CO-IP-MS. b The Venn diagram showed the number of significant upregulated (black) and downregulated (blue) proteins identified in response to TRIM31 knockdown, and the protein expression fold change is MCF7-1/MCF7-NC > 1.5 or <0.67, and unique peptide ≧ 2 is defined as a significantly different protein. TRIM31 interaction candidates were identified in the Co-IP process (red). The below showed potential substrates of the TRIM31. c, d The HEK293 cells were co-transfected with Flag-TRIM31 and HA-P53 plasmids for 48 h, and then the cells were treated with MG132 (20 μM) for 4 h. The lysates of cells were immunoprecipitated with Flag-tag antibody and then western blot assay with HA-tag antibody (c); immunoprecipitated with HA-tag antibody and immunoblotted with Flag-tag antibody (d). e The MCF7 cell lysates were immunoprecipitated with Ig G and anti-p53 antibody and the expression of TRIM31 and p53 were detected by western blot. f GST-pull down was performed to verify the direct interaction of TRIM31 and P53 and the components were detected by the western blot assay with anti-GST and anti-His antibody. g The schematic diagram showed structural domains of TRIM31 and p53 protein. h HEK293 cells were transfected with HA-P53 and Falg-TRIM31 or several TRIM31 deletion mutants. The whole-cell lysates were immunoprecipitated with anti-Flag beads and immunoblotted with anti-Flag and anti-HA antibodies. i HEK293 cells were co-transfected with Falg-TRIM31 and HA-P53 or several P53 deletion mutants. The whole-cell lysates were immunoprecipitated with anti-HA antibodies and immunoblotted with anti-Flag and anti-HA antibodies.