AtELF3β promotes flowering by interacting with AtELF3α to inhibit the formation of the evening complex. A) The indicated plasmids were co-transformed into yeast strain Y2HGold. Interactions between bait and prey were examined on synthetic defined (SD) medium-LW and selective medium (SD-LWHA). B) Firefly luciferase complementation imaging (LCI) assays in N. benthamiana leaves showing that AtELF3β can interact with AtELF3α. The pseudocolor bar indicates the range of luminescence intensity. C) AtELF3β interacts with and co-immunoprecipitates with AtELF3α in vivo. N. benthamiana leaves were infiltrated with 35S:GFP-AtELF3β and 35S:AtELF3α-FLAG constructs via Agrobacterium-mediated infiltration. AtELF3α-FLAG was detected in the immunoprecipitated GFP–AtELF3β complex. D) The interaction between AtELF3α and LUX decreases in the presence of AtELF3β in the LCI assays. N. benthamiana leaves were co-infiltrated with four sets of constructs at symmetric positions with equal amounts of Agrobacteria. The pseudocolor bar indicates the range of luminescence intensity. E) The interaction between AtELF3α and LUX decreases in the presence of AtELF3β in an in vivo co-immunoprecipitation. AtELF3α-FLAG was detected in the immunoprecipitated GFP–LUX complex, indicating that there is a physical association between LUX and AtELF3α in vivo. With the addition of HA-AtELF3β expression, AtELF3α-FLAG was not detected in the immunoprecipitated GFP–LUX complex. The asterisk indicates the GFP–LUX band.