Fig. 2.

Effects of JAB1 inhibition on Cul3 neddylation and substrate protein abundance. Myc-tagged KLHL3 or WNK4 was cotransfected into HEK293 cells with either JAB1 siRNA or control siRNA. The proteins were examined by immunoblot in cells with endogenous WT Cul3. JAB1 siRNA decreased JAB1, KLHL3, and WNK4 abundance and increased NEDD8 abundance and the neddylated form of Cul3 (top band). β-actin was used as a loading control. Con, control; Cul3, cullin 3; HEK, human embryonic kidney; JAB1, jun activation domain-binding protein-1; KLHL3, kelch-like 3; NEDD8, neuronal precursor cell expressed developmentally downregulated protein 8; WNK, with-no-lysine kinase; WT, wild type.