Fig. 2

Effect of B-box mutations on CO homodimerization. A. Schematic illustration of co mutant alleles. Sequences of the N-terminal regions of the co mutant alleles co-2, co-3, co-4, and co-6 were amplified using the mRNA extracted from each mutant allele and introduced into the pDEST22 and pDEST32 vectors. B. Yeast two-hybrid assays coupled with β-galactosidase measurement. C. In vitro binding assays. Full-length co mutant cDNA was introduced into the pDEST15 and pDEST17 vectors to generate the GST and 6XHis fusion proteins, respectively. Presence or absence of the interactions was confirmed through western blotting against anti-GST and 6XHis antibodies