Fig. 3

TRIM59 promotes ABHD5 protein degradation through ubiquitination. a. Proteasome inhibitor MG132 inhibited the downregulation of ABHD5 protein levels induced by HA-TRIM59 expression in HEK-293 cells. HEK-293 cells were transfected with HA-TRIM59 or HA-TRIM59-∆RING vector. The indicated proteins were measured by western blotting. b. HA-TRIM59 expression decreased ABHD5 protein half-life in cells. HEK-293 cells with ectopic HA-TRIM59 expression and control cells were transfected with the Myc-ABHD5 vector. The cells were treated with 50 μg/ml CHX for indicated time periods before being collected for western blot assays. c. Knockdown of endogenous TRIM59 increased Myc-ABHD5 protein half-life in THP-1 macrophages. d. The effects of expression of HA-TRIM59 and its mutants on ubiquitination of Myc-ABHD5 in HEK-293 cells analyzed by in vivo ubiquitination assays. e. Knockdown of endogenous TRIM59 decreased the ubiquitination of Myc-ABHD5 in THP-1 macrophages analyzed by in vivo ubiquitination assays. f. Mutations of TRIM59 that impaired TRIM59’s ubiquitination activity impaired the ability of HA-TRIM59 to degrade Myc-ABHD5 protein in THP-1 macrophages