DCAF11 levels drastically reduced in all csn mutants, possibly because of rapid autoubiquitination-mediated degradation.

(A) Immunoprecipitation assay confirming the interaction between Flag-Cul4 and Myc-DCAF11. Wild-type strain and wild-type strain expressing Myc-DCAF11 were used as negative controls. (B) Western blot analyses showing the degradation of Myc-DCAF11 in csn mutants after the addition of CHX. (C) Amino acid alignment of conserved WDXR motifs of DCAF11 homologs. (D) Immunoprecipitation assay with c-Myc antibody showing that Myc-DCAF11R432A failed to interact with Flag-DDB1. (E) Myc immunoprecipitation assay showing that Myc-DCAF11R432A failed to interact with Flag-Cul4. (F) Degradation of Myc-DCAF11R432A expressed in the wild-type strain and all csn mutants after the addition of CHX. Densitometric analyses from four independent experiments showing degradation of DCAF11 (G) and DCAF11R432A (H).