Figure 5.

GhFP2 interacts with GhACE1. A, Y2H assay of the interaction between GhFP2 and GhACE1. pGBKT7 and pGADT7 empty vectors were used as controls. B, LCI assay of the interaction between GhFP2 and GhACE1. GhFP2 was fused to the amino-terminal of firefly luciferase (LUCn), and GhACE1 was fused to carboxyl-terminal of firefly luciferase (LUCc), respectively. The LUCc-GhACE1 and GhFP2-LUCn constructs were transiently co-expressed in leaves of N. benthamiana, using LUCn and LUCc as the controls. Fluorescence signal intensities represent their binding activities. Right bars indicate heat map' scales of the signal intensity. C, CoIP of transiently co-expressed GhFP2-HA and GFP-GhACE1 in N. benthamiana leaves. Soluble protein extracts before (input) and after (IP) immunoprecipitation with anti-GFP antibody-conjugated beads were detected by immunoblot with anti-HA antibody.