Fig. 5.

Effects of proteasome inhibition on Cul3Δ403–459-mediated KLHL3 degradation. The pathway for degradation of KLHL3 by the Cul3Δ403–459 mutant was examined by inhibiting the proteasomal pathway with the drug MG132. HEK293 cells were cotransfected with myc-KLHL3 and either no Cul3 or FLAG-Cul3Δ403–459. The cells were incubated with vehicle or 10 μM MG132 for 18 h before harvesting. Immunoblot analysis showed that inhibition of the proteasomal pathway partially blocked Cul3Δ403–459-mediated KLHL3 degradation. Right, quantitative analysis of KLHL3 protein abundance. GAPDH was used as a loading control. Data represent individual values as well as means ± SE relative to control. Statistical differences were examined by one-way ANOVA with Tukey’s post hoc analysis. Con, control; Cul3, cullin 3; HEK, human embryonic kidney; KLHL3, kelch-like 3; NS, not significant; Veh, vehicle.