Fig. 3: FHY3 and FAR1 directly interact with SPL9 and SPL15.
From: Arabidopsis FHY3 and FAR1 integrate light and strigolactone signaling to regulate branching
a Yeast two-hybrid assay shows that both SPL9 and SPL15 interact with FHY3 and FAR1. LexA and JG serve as the negative controls. b BiFC assay shows that both SPL9 and SPL15 interact with FHY3 and FAR1 in N. benthamiana leaf epidermal cells. Both FHY3 and FAR1 were fused to the N-terminal fragment of YFP (nYFP) and both SPL9 and SPL15 were fused to the C-terminal fragment of YFP (cYFP). The AT-HOOK-RFP marker was used to indicate the nuclei. The interaction between nYFP and SPL9-cYPF or SPL15-cYPF serves as the negative controls. Scale bar = 50 μm. c Pull-down assay shows that FHY3 directly interacts with both SPL9 and SPL15 in vitro. GST-SPL9SBP or GST-SPL15SBP proteins were incubated with protein extracts containing His-FHY3-I and further immobilized with glutathione sepharose beads. d Co-IP analysis shows the interaction between FHY3/FAR1 and SLP9/15 using a transient expression system in tobacco leaves. Immunoprecipitation was performed using anti-HA matrix and the co-immunoprecipitated proteins were detected with anti-MYC antibodies. This assay was repeated for three times and similar results were obtained.
