Fig. 7. TRIM7 interacts with DUSP10 and facilitates degradation via K63-linked ubiquitination.

a Schematic of the design for IP Flag-TRIM7 and subsequent LC-MS/MS analysis. b Co-IP assay to examine the interaction between endogenic TRIM7 and DUSP10 proteins. c Schematic representation of full-length (FL) TRIM7 and DUSP10, as well as their truncated mutant constructs. d, e IP mapping analysis of the interaction domains between TRIM7 and DUSP10 in HEK293T cells co-transfected with plasmids encoding HA-DUSP10 and Flag-TRIM7 or its truncated mutants (d); or plasmids encoding Flag-TRIM7 and HA-DUSP10 or its truncated mutants (e). f, g Ubiquitination of DUSP10 in HEK293T cells co-transfected with Myc-Ub, HA-DUSP10, and Flag-tagged TRIM7 wildtype or its E3 dead ligase mutant plasmids (f) or TRIM7 knockdown plasmids (g). h Ubiquitination screening to identify the ubiquitin type of DUSP10 catalyzed by TRIM7 in HEK293T cells co-transfected with HA-DUSP10, Flag-TRIM7, and the indicated Myc-Ub constructs. i Ubiquitination analysis of DUSP10-WT and DUSP10-K425R in 293T cells transfected with or without Flag-TRIM7 plasmids. j Immunoblotting analysis of DUSP10 expression in 293T cells co-transfected with HA-DUSP10 and different dose of Flag-tagged TRIM7 wildtype or TRIM7-DL plasmids. k Immunoblotting (left panel) and quantification (right panel) of DUSP10 protein levels to assess the effect of TRIM7-WT or TRIM7-DL on DUSP10 half-life via CHX chase assay (n = 3; P values determined by one-way ANOVA). Data are presented as mean ± SD. l Western blot analysis of DUSP10 expression in 293T cells transfected with the indicated plasmids, followed by treatment with CHX alone or in combination with MG132, CHQ or NH₄Cl prior to cell harvest. m Western Blot analysis of DUSP10 and TRIM7 protein expression in liver tissues from mice fed a HFD (left panel) or HFFC (right panel) diet (n = 4). These data were obtained from at least three independent experiments using independent samples. Source data are provided as a Source Data file.