Skip to main content
Advertisement
Browse Subject Areas
?

Click through the PLOS taxonomy to find articles in your field.

For more information about PLOS Subject Areas, click here.

< Back to Article

Mechanism of Cullin3 E3 Ubiquitin Ligase Dimerization

Figure 2

Cul3-Cul3 binding requires the Cul3 N-terminus but not the Cul3 C-terminus.

A, The cells were cotransfected with Cul3-HA and Cul3(1–427)-V5 (ΔCT), as indicated at the top of each panel. The cells were lysed, and the lysates were subjected to V5 immunoprecipitation (IP), Immunoprecipitates and aliquots of the cell lysates were analyzed by Western blotting with the indicated antibodies. B, cells were transfected with wild type (wt) Cul3-V5, Cul3(CT)-V5 and Cul2(NT)-Cul3(CT)-V5, as indicated. Two days after transfection, the cells were treated with 25 ìM MG-132 for 6 h, where indicated, and then lysed, followed by Western blotting of cell lysates with V5 antibody. C, HEK293 cells were transfected in 60-mm cell culture plates for 2 days with expression constructs for the proteins as indicated at the top of each panel. After cell lysis, the cell lysates were subjected to V5-immunoprecipitation and immunoblotting with V5 and HA antibodies.

Figure 2

doi: https://doi.org/10.1371/journal.pone.0041350.g002