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. 2004 Apr 15;18(8):856–861. doi: 10.1101/gad.1177904

Figure 3.

Figure 3.

Point mutations in RhoBTB2 and Cul3 disrupt their interaction. (A) A Y284D mutation in RhoBTB2 disrupts the interaction with Cul3. 293T cells were transfected with the indicated plasmids and anti-HA immune complexes isolated and analyzed sequentially for the presence of RhoBTB2 and Cul3 by Western blotting. The Y284D RhoBTB2 mutant was consistently expressed at levels three to fivefold higher than wild type; consequently, four times less RhoBTB2-Y284D immunoprecipitate than wild type was loaded to allow easier assessment of the relative amounts of Cul3. (B) A Y58K mutation in Cul3 disrupts the interaction with RhoBTB2. 293T cells were transfected with the indicated plasmids and anti-HA immune complexes isolated and analyzed sequentially for the presence of RhoBTB2 and Cul3 by Western blotting.