Fig. 6

TRIM3 is localized in the cytosol and interact with P53 in breast cancer cells. a Intracellular localization analysis of P53 and TRIM3 by immunofluorescence assay. MCF-7 cells were cultured in normal medium before fixation. Intracellular localization of TRIM3 (green) and P53 (red) were shown. Nuclei (blue) were stained with 4′,6-diamidino-2-phenylindole (DAPI). b Co-IP assay revealed association between endogenous P53 and TRIM3 protein in MCF-7 cells. MCF-7 cells were harvested with RIPA lysis buffer. CO-IP was performed using antibody as indicated. c P53 domain structure and deletion variants in the study. d TRIM3 domain structure and deletion variants in the study. e P53 DNA binding domain is required to mediate the TRIM3-P53 interaction. HEK293 cells were transfected with myc-TRIM3 together with GFP-P53 full length or variants. After 24 h, cells were harvested with NP-40 lysis buffer. CoIP was performed and the possible interacted P53 variants were detected by GFP antibody. f RING domain of TRIM3 is required to mediate the TRIM3-P53 interaction. HEK293 cells were transfected with Flag-P53 together with GFP-TRIM3 full length or variants. After 24 h, cells were harvested with NP-40 lysis buffer. CoIP was performed and the possible interacted TRIM3 variants were detected by GFP antibody