(A) Dual-membrane yeast two-hybrid assays showing interaction of LLG2/3 with ectodomains of BUPS1/2 and ANX1/2. Yeast cells containing indicated plasmids were plated on the growth medium (-Leu-Trp) or on selection medium (-Leu-Trp-His-Ade).
(B and C) Interactions between LLG2/3 with BUPS1/2 (B) and ANX1/2 (C) ectodomains by pull-down assays. LLG2/3 were expressed in E. coli, and ectodomains of BUPS1/2 and ANX1/2 were transiently generated in tobacco leaves. Pull-down assays were performed with Ni Sepharose, and western blots were probed with antibodies against FLAG and His (α-FLAG or α-His), respectively.
Similar results were obtained by three independent experiments.