Extended Data Fig. 7 |. BSL1 Physically Interacts with YDA in vitro and in vivo.

a, In vitro pull-down assays using purified recombinant proteins show GST-BSL1 interaction with MBP-YDA. MBP-YDA used as bait. GST (negative control) or GST-BSL1 interaction were detected by anti-GST. b, Co-IP data show in vivo interaction between BSL1 and YDA. BSL1-FLAG was used as bait to detect the binding of YDA^KI-Myc in 5-dpg Arabidopsis seedlings. The expression of BSL1-FLAG is driven by the TMM promoter and the expression of YDA is driven by the BASL promoter. YDA^KI (Kinase Inactive YDA) was used to avoid the activity of YDA strongly suppressing plant growth. Data represent results of experiments repeated three times in a and b. c, BiFC assays show the interaction between BSL1 and YDA^KI (kinase inactive YDA variant used to suppress active YDA-triggered cell death) occurs at the cell membrane in N. benthamiana leaf epidermis. Positive protein-protein interactions were visualized by YFP signal (yellow). Half YFPs (nYFP or cYFP) were uses as negative controls. Scale bars, 50 μm.