Fig. 5.

TRIM11 increases the mono-ubiquitinated ERα while decreases K11-, K27- and K48-dependent polyubiquitination on ERα protein. (A-I). HEK293 cells were transfected with 2 µg Flag- ERα plasmid, 0.5 µg Myc-TRIM11 plasmids, and 0.5 µg HA-K6, -K11, -K27, -K29, -K33, -K48, -K63, -K0 or -Ub plasmids. The cell extracts were immunoprecipitated with HA antibody. The specific polyubiquitinated and mono-ubiquitinated ERα were detected via western blotting analysis. (J). TRIM11 depletion decreases ERα mono-ubiquitination in MCF7 cells. MCF7 cells were transfected with siTRIM11 or siControl. After 48 h, cells were treated with 10 µM MG132 for 6 h, cell lysates were prepared for western blot analysis and anti-HA antibody was used to detect mono-ubiquitylated ERα.