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. 2014 Jul 10;15:28. doi: 10.1186/1471-2121-15-28

Figure 2.

Figure 2

The newly identified Cul3 binding proteins (CLWs) bind to Cul3 in cells. HEK293 cells were transfected with vectors expressing FLAG-tagged Cul3 and HA-tagged CLWs. A. Western blot analysis shows expression and binding of Cul3 and two LRR domain-containing proteins, LRR1 (left two lanes) and LRR5 (right two lanes). B. Western blot analysis shows expression and binding of Cul3 and two WD40 domain-containing proteins, W14 (left two lanes) and W16 (right two lanes). C. Wild type Ctb73 was cotransfected with wild type Cul3 (first lane), or a Cul3Δ51-67 mutant (second lane) or expressed alone (third lane). Upper panel shows relative binding between Cul3 and Ctb73, while lower panel shows the expression levels of Ctb73. D. Immunoprecipitation assays to detect binding between two different LRR proteins: LRR1 (lane 1–3) and LRR5 (lane 4–6). Wild type Cul3 was expressed in lanes 1 and 4, while the Cul3Δ51-67 mutant was expressed in lane 2 and 5. Lastly, Cul3 was not transfected in lane 3 and 6, which serves as negative controls for the immunoprecipitation assays. See also, Additional file 4: Table S4.