Figure 7. Co-inactivation of CAND-1 and neddylation pathway components.
(A) Whole-worm lysates from wild type and cand-1 mutants with or without RNAi depletion of csn-5 (COP9 Signalosome deneddylase) or ubc-12 (Nedd8 E2), and probed with anti-CUL-2 and anti-tubulin antibodies. (B) Quantitation of the neddylated and unneddylated CUL-2 isoform levels in (A) relative to tubulin levels. The levels of CUL-2 isoforms in cand-1 mutants are normalized to the total CUL-2 signal of wild-type animals (set to 100 a.u.); and the percentage of neddylated isoform within a genotype is provided in white lettering. (C) Total-worm lysates prepared from wild type and cand-1 mutants expressing CUL-4∷FLAG with or without csn-5 or ubc-12 RNAi, and probed with anti-FLAG or anti-tubulin antibodies. (D) Quantitation of slower-migrating covalently-modified and unmodified CUL-4∷FLAG isoforms in (C). Normalization and labeling are as in (B).