Extended Data Fig. 3: The interaction between NSs and AtCOI1, AtTIR1, AtMAX2 or Tsw-PRL.

a, Co-immunoprecipitation (Co-IP) analysis of the interaction between NSs and AtCOI1/AtTIR1/AtMAX2. YFP-AtCOI1, YFP-AtTIR1 and YFP-AtMAX2 were used to Co-IP the NSs-FLAG inN. benthamiana工厂。YFP是使用d as a control. The upper arrow indicates the band of YFP-AtCOI1, YFP-AtTIR1 and YFP-AtMAX2 protein; the lower arrow indicates the band of YFP protein. Blots were detected using YFP and FLAG specific antibodies. Ponceau S staining was used to estimate sample loading.b, SLC analysis of the interaction between NSs and AtCOI1/AtTIR1/AtMAX2in planta. cLUC-AtCOI1, cLUC-AtTIR1, cLUC-AtMAX2 or cLUC control vector were co-expressed with nLUC-NSs or nLUC control vector inN. benthamianaplant leaves. Luciferase activity was detected at 48 hpi.c, BiFC analysis of the interaction between NSs and AtCOI1/AtTIR1/AtMAX2 or Tsw-PRLin planta. nYFP-AtCOI1, nYFP-AtTIR1, nYFP-AtMAX2, nYFP-Tsw-PRL or nYFP control vector was co-expressed with cYFP-NSs or cYFP control vector inN. benthamianaplant leaves. The reconstituted YFP fluorescence signals were examined by confocal microscopy and photographed at 48 hpi. Scale bar, 50 µm.d, NSs interacts with AtCOI1/AtTIR1/AtMAX2 in the nucleus assayed by BiFC. cYFP-NSs and nYFP-AtCOI1, nYFP-AtTIR1 or nYFP-AtMAX2 were co-expressed with H2B-mCherry, a nuclear marker, inN. benthamianaplant leaves. The white arrow indicates the co-localization of NSs, AtCOI1/AtTIR1/AtMAX2 and H2B in the nucleus. Scale bar, 50 µm.e, Co-IP analysis of the interaction between NSs and Tsw-PRLin planta. YFP-Tsw-PRL was used to Co-IP NSs-FLAG inN. benthamiana工厂。YFP是使用d as a control. Blots were detected using YFP and FLAG specific antibodies. Ponceau S staining is used to estimate sample loading.f, SLC analysis of the interaction between NSs and Tsw-PRLin planta. cLUC-Tsw-PRL or cLUC control vector was co-expressed with nLUC-NSs or nLUC control vector inN. benthamianaplant leaves. Luciferase activity was detected at 48 hpi.g, Y2H analysis of the interaction between NSs and AtCOI1, AtTIR1, AtMAX2 or Tsw-PRL. pGBKT7 empty vector containing a DNA binding domain (BD) and pGADT7 empty vector containing an activation domain (AD) were used as negative controls. BD-NSs and AD-NSs were used as a positive control. The yeast co-transformed with BD and AD derivative constructs was plated and assayed on both SD/-L-T (left) and SD/-L-T-H-A (right) dropout media. Experiments were repeated at least three times with similar results.