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Fig. 6. The CCT domain of CO is necessary for the in vitro interaction with
SPA1. (A) Schematic representation of the deletion-derivatives of
CO or His6-CO used in the in vitro binding assay. (B)
Mapping of the SPA1-interacting domain of CO. Recombinant
35S-labelled CO or indicated CO-deletion proteins were incubated
with partially 35S-labelled GAD-SPA1 or GAD, respectively, and
co-immunoprecipitated with anti-GAD antibodies. Supernatant fractions (1.6%)
and 33.3% of the pellet fractions were resolved by SDS-PAGE and visualized by
autoradiography using a phosphorimager. (C) Quantification of the
fractions of prey proteins that were co-immunoprecipitated by the indicated
bait proteins GAD-SPA1 or GAD. Error bars depict the s.e.m. from two replicate
experiments.