Fig. 2

TbCSB βC1 interacts with NbGeBP in vitro and in vivo. a Alignment of GeBP protein sequences from S. lycopersicum, S. tuberosum, N. benthamiana, C. annuum, A. thaliana, and T. aestivum. b Phylogenetic tree of GeBP was constructed by the Maximum-Likelihood method using the protein sequences from S. lycopersicum, S. tuberosum, N. benthamiana, C. annuum, A. thaliana, and T. aestivum. c Y2H assay of the interaction between the βC1 and NbGeBP in yeast cells. The transformed yeast cells were subjected to tenfold serial dilutions and plated on SD/-Trp-Leu-His-Ade medium for 3 d. Yeast cells co-transformed with AD-T and BK-P53 were served as positive controls; yeast cells co-transformed with AD-T7 and BK-Lam were used as negative controls. d Y2H assay of the interaction between the βC1 and CaGeBP and StGeBP in yeast cells. e Y2H assay of the interaction between the βC1 and AtGeBP in yeast cells. f LCI analysis of the interaction between βC1 and NbGeBP in N. benthamiana leaves. βC1-NLuc co-expressed with CLuc or NLuc co-expressed with CLuc-NbGeBP, and NLuc co-expressed with CLuc, were used as negative controls. g BiFC analysis of the interaction between NbGeBP and βC1 in N. benthamiana leaves. The RFP-H2B is nuclear localization marker. Co-expression of Y^N-NbGeBP and Y^C, or Y^N and Y^C-βC1 were used as negative controls. Confocal imaging was performed at 48 hpai. Scale bars: 50 μm. h BiFC analysis of the interaction between AtGeBP and βC1 in N. benthamiana leaves. Co-expression of Y^N-SlGeBP and Y^C-βC1 were used as positive controls, and co-expression of Y^N-βC1 and Y^C, or Y^N and Y^C-AtGeBP were used as negative controls. Confocal imaging was performed at 48 hpai. Scale bars: 100 μm. i Y2H assay of the interaction between the βC1 and NbGeBP-deletion derivatives (△1–7aa, △16–23aa, △36–45aa, △46–138aa, and △338–377aa) in yeast cells. These experiments were performed with three independent biological replicates with similar results