Fig. 4: TRIM25 regulates the stability of the BRD7 protein by regulating K48-linked ubiquitination.

A Western blot analysis of BRD7 protein levels in MDA-MB-231 and MCF7 cells were transfected with siNC, siTRIM25#1, or siTRIM25#2. B BC cells transfected with siNC or siTRIM25 incubated with CHX (50 μM) for 0, 1, 2, and 4 h, the BRD7 protein level was detected by western blot. C BC cells were transfected with the TRIM25 plasmid or the vector plasmid and further incubated with MG132 (20 μM) for 4 h, the BRD7 protein level was detected by western blot. D MDA-MB-231 and MCF7 cells were co-transfected with siNC, siTRIM25#1, or siTRIM25#2 together with HA-Ub, after MG132 (20 μM, 4 h) treatment, the ubiquitination level of BRD7 was detected via IP and western blot analysis. E Co-IP and ubiquitination experiment on the changes in BRD7 ubiquitination levels after cotransfection with TRIM25 plasmid and K48-Ub or K63-Ub plasmid in MCF7 cells. F Co-IP and ubiquitination experiment was performed to detect the ubiquitination levels of BRD7 in MCF7 cells co-transfected with Flag-TRIM25 plasmid or Flag-TRIM25ΔRING plasmid and HA-UB plasmid.