Figure 5.

The eta3 Mutation Does Not Alter SCF^TIR1 Assembly or Substrate Binding.

(A) Anti-c-myc antibody was used to immunoprecipitate the SCF^TIR1 complex from extracts prepared from tir1-1 and eta3 seedlings expressing a TIR1-myc fusion gene. Immunoprecipitates (IP) were immunoblotted with anti-ASK1 and anti-AtCUL1 antibodies as described previously (Gray et al., 1999).

(B) Recombinant GST-AXR3 protein was used to pull down the SCF^TIR1 complex from tir1-1[TIR1-myc] and eta3[TIR1-myc] seedling extracts. Pull downs were immunoblotted with anti-c-myc antibody to detect the presence of the TIR1-myc fusion protein.