Extended Data Fig. 4: Yeast two hybrid screening identified AtTCP21 as a common interactor for NSs, AtCOI1, AtTIR1, AtMAX2 and Tsw-PRL.

a, Interaction of full-length AtTCP7 and AtTCP14 with NSs in yeast. AD, pGADT7 vector. BD, pGBKT7 vector. The yeast co-transformed with BD and AD derivative constructs was plated on both SD/-L-T (left) and SD/-L-T-H-A (right) dropout media. b, Number and frequency of AtTCP7 and AtTCP14 identified in Y2H library screening using NSs as bait. c, Y2H analysis of the interaction between NSs and 24 TCP family members. The full-length coding sequence of 24 TCP family members from Arabidopsis plant was constructed into AD vector and co-transformed with BD-NSs or BD control vector into yeast and their growth and interaction were assayed on both SD/-L-T and SD/-L-T-H-A dropout media. d, Phylogenetic tree analysis of 24 TCP family members from Arabidopsis. The phylogenetic tree was constructed using MEGA 7.0 by the neighbor-joining method and bootstrap values (1000 replicates) are shown on the branches. e, Y2H assay showing direct interaction between AtTCP7/14/17/21 and AtCOI1/AtTIR1/AtMAX2 or between AtTCP7/14/17/21 and NSs. The results for SD/-L-T dropout media are shown in this Figure and results for SD/-L-T-H-A dropout media are shown in Fig. 2a. f, BD-AtCOI1, BD-F-box (AtCOI1), BD-LRR (AtCOI1) or BD vector control were tested for the interaction with AD-AtTCP21 or AD vector control in yeast. g, AD-AtTCP21, AD-AtTCP21 (1-103), AD-AtTCP21 (104-240), AD-AtTCP21 (30-103), AD-AtTCP21 (30-136), AD-AtTCP21 (137-240) or AD vector control were tested for the interaction with BD-AtCOI1 or BD vector control in yeast. The schematic diagram of truncation mutants of AtTCP21 was shown above the Y2H assays. h, SLC analysis of the interaction between NSs and AtTCP21 in planta. cLUC-AtTCP21 or cLUC was co-expressed with nLUC-NSs or nLUC in N. benthamiana plant leaves. Luciferase activity was detected at 48 hpi. i, Y2H analysis of the interaction between AtTCP21 and LRR domain of different proteins. BD-FBL6-LRR, BD-FBL17-LRR, BD-RPM1-LRR, BD-HRT-LRR, BD-FLS2-LRR, BD-EFR-LRR or BD vector control were tested for the interaction with AD-AtTCP21 or AD vector control in yeast. AD, pGADT7 vector. BD, pGBKT7 vector. FBL6 and FBL17 are two F-box family proteins; RPM1 and HRT are two CC-NLRs; FLS2 and EFR are two PRRs. The yeast cotransformed with BD and AD derivative constructs were plated and assayed on both synthetic double dropout media -Leu-Trp (SD/-L-T) and synthetic quadruple dropout media -Leu-Trp-HIS-Ade (SD/-L-T-H-A).