Fig. 1

Yeast two-hybrid assay to isolate CO-binding proteins. a Schematic representation of the CO deletion derivative, COΔ133-174. b Gal4 DNA binding domain (BD) fusions of CO and COΔ133-174 were expressed in yeast. Different concentrations of 3-AT (0.1, 0.5, 1, 5 mM) and Adenine (Ade) were used to validate elimination of the transcriptional activity for the reporter genes in the COΔ133-174 protein-expressing yeast cells. c After isolating FHTH1 as a CO-interacting protein in the yeast two-hybrid screen, co-transformation and serial dilution assays using the BD-COΔ133-174 and AD-FHTH1 constructs were performed to confirm the protein–protein interaction. Yeast cells were grown on SD agar medium lacking Trp and Leu (left panel) or lacking Trp, Leu, His, and Ade (right panel). The HY5-STO interaction was included as a Ref. [37]