Fig. 4: HAUSP removes ubiquitin chains from LZTS2 to stabilize it.
From: SPOP and HAUSP bidirectionally regulate LZTS2 ubiquitination to modulate the Wnt pathway
A HAUSP increased Myc-LZTS2 protein in a dose-dependent manner in 293 T cells. B Knockdown of HAUSP decreased endogenous LZTS2, YAP and GLI3 protein levels in HCT-116 cells. C 293 T cells were treated with CHX (20 μg/ml) for the indicated times. Of note, knockdown of HAUSP promoted the degradation of endogenous LZTS2. D The deubiquitinase-deficiency mutant HAUSP-CA failed to increase Myc-LZTS2 protein. E HAUSP enhanced endogenous LZTS2 protein in a dose-dependent manner in HT-29 cells, which was abolished by treatment with 10 μM of HAUSP inhibitor P22077 for 24 h. F Treatment with three HAUSP inhibitors (P5091, USP7-IN-1, or P22077) at the indicated concentrations for 24 h apparently downregulated endogenous LZTS2 protein in HCT-116 cells. G Immunoblots of immunoprecipitates (top two panels) or whole cell extracts (bottom three panels) from 293 T cells transfected with the indicated plasmids. HAUSP suppressed K48- and K63-linked polyubiquitination on LZTS2. Above all, Actin acts as a loading control.
