Fig. 6.

GhWRKY27 directly targets the promoters of GhCYP94C1 and GhRipen2–2. a Interaction between GhWRKY27 and the candidate target genes GhASPG1, GhWRKY1, GhRipen2–1, Gh20ox2, GhCYP94C1, GhGH3.5, GhIAA15A, and GhRipen2–2 in Y187 yeast cells. The binding ability of GhWRKY27 with the target promoter sequences was identified based on the growth status of transformed Y187 yeast cells on SD-TL and SD-TLH + 200 mM 3-AT medium. The combination of pGADT7-GhWRKY27 and pHIS2 was used as a negative control. b Wild and mutated promoter probe sequences of GhCYP94C1 were used in the EMSA assay. The W-box sequence 5′-TGACT-3′ was mutated to 5′-TGAAA-3′. c Wild and mutated promoter probe sequences of GhRipen2–2 were used in the EMSA assay. The W-box sequence 5′-TTGACT-3′ was mutated to 5′-TTGAAA-3′. d GhWRKY27 binds to the W-boxes in the promoter of GhCYP94C1 in the EMSA assay. e GhWRKY27 binds to the W-boxes in the promoter of GhRipen2–2 in the EMSA assay. A monoclonal antibody against GhWRKY27 (anti-GhWRKY27) was employed in the supershift assay. “–” indicates absence, while “+” indicates presence