Yeast two-hybrid assay

The full-length coding sequences of AtCLV2, AtCLV1, AtCRN and AtCLV1-ECD were each fused to the Gal4 DNA binding domain of pGBKT7. The full-length coding sequences of AtCLV3 and GhCLE5 were each fused to the Gal4 activation domain in pGADT7. The constructed bait and prey composition was co-transformed into the yeast strain gold yeast. Two days after growth on SD-Leu/-Trp plates, the interaction between the bait and the prey was observed on the SD-Leu/-Trp/-His selective medium. Yeast strains containing pGBKT7_AtCLV3 and pGBKT7_GhCLE5 and a negative pGADT7 vector were used as negative controls. The constructed bait and prey composition was co-transformed into the yeast strain gold yeast following the kit manufacturer’s instructions (Frozen-EZ Yeast Transformation II Kit, Zymo Research). All primer information can be found in Supplemental Table ⁶.