Fig. 4.

TRIM3 associates with ER alpha and modulates ER alpha stability. a Intracellular localization of ER alpha (red) and TRIM3 (green) is shown. Nuclei (blue) were stained with 4′,6-diamidino-2-phenylindole (DAPI) in MCF-7 cells. b Co-IP was performed using antibody as indicated. c ER alpha protein levels were determined by western blotting. HEK293 cells were transfected with 1 μg ER alpha together with different amounts of TRIM3 plasmids. d ER alpha protein levels were determined by western blotting. Actin was used as internal control. HEK293 cells were transfected with TRIM3 and ER alpha plasmids. After 24 h, cells were treated with 10 μM MG132 or vehicle for 6 h. e and f ER alpha protein levels were determined by western blotting. Actin was used as internal control. HEK293 cells were transfected with HA-ERα plasmid and Myc-tagged or Myc-TRIM3 plasmids. After 24 h, cells were treated with 100 µM cycloheximide (CHX) for the indicated times. The results are representative of three independent experiments. The relative density of ER alpha was measured by ImageJ software