Fig 6.

Coimmunoprecipitation assay. Protein A was coexpressed with either wt CP or its indicated mutants in N. benthamiana leaves by agroinfiltration. (A) Expression of protein A and CP in total protein extracts was confirmed by Western blotting with anti-protein A and anti-CP antibodies, respectively. Total protein extracts from leaves infiltrated with the empty vector or CP only were used as controls. (B) For coimmunoprecipitation, total protein extracts of each sample were incubated first with anti-protein A antibodies, followed by complex precipitation with protein G-agarose. The resulting coimmunoprecipitated products were subjected to Western blotting with anti-protein A and anti-CP antibodies. The effect of RNase A treatment on coimmunoprecipitated products was verified by Western blot analysis as described above.