Figure 2

No mutant BRCA2 protein was detected in overexpression experiments. (a) Lysates of transfected COS7 cells were subjected to electrophoresis through 6–18% gradient gel and blotted with an antibody against the V5 tag (α-V5): 1, untransfected cells; 2, pIND-LacZ transfected cells; 3, cells cotransfected with both pIND-LacZ and pIND-BRCA2 999del5; and 4, cells transfected with pIND-BRCA2 999del5. (b) Nickel-coated agarose beads were used to adsorb recombinant proteins under native conditions. The proteins were subjected to electrophoresis through 6–18% gradient gel and blotted with α-V5: 1, untransfected COS7 cells; 2, cells overexpressing pIND-BRCA2 999del5; and 3, cells overexpressing pIND-LacZ. (c) α-V5 was used to precipitate recombinant proteins and for immunoblotting: 1, untransfected COS7 cells; 2, cells transfected with pIND-LacZ; 3, cells transfected with pIND-BRCA2 999del5 without induction; and 4, cells transfected with pIND-BRCA2 999del5 with induction.