Extended Data Fig. 1: Proxitome profiling of the BSR1 protein.
a, Diagram of the constructs used for the identification of proximal proteins to BSR1. The full-length BSR1 protein and its CC, NB-ARC, and LRR domains are shown on the top. b, Functional validation of TurboID-fused BSR1 protein. Fusion of TurboID to the C-terminus of BSR1 has no discernible effect on the ability of BSR1 to induce cell death. Cell death phenotype under bright light (left) or after trypan blue staining (right) is shown. c, Workflow for the PL analysis in this study. Agrobacterium mixtures containing TurboID fusions, with (+TGB1) or without TGB1 (−TGB1) under the control of the 35S promoter, were co-infiltrated into the leaves of N. benthamiana plants. At 36 hpi, 200 µM biotin was infiltrated into pre-infiltrated leaves. 8 h later, the infiltrated leaves were harvested for subsequent processing as indicated on the panel. Each treatment was performed with three independent biological replicates (n = 3 plants for each technical replicate). The numbers 126 to 131 above the leaf schematic indicate the 6-plex TMT reporter ion masses at m/z 126 to 131. d, e, Immunoblot analysis to confirm the enrichment of biotinylated proteins without (d) and with (e) the addition of TGB1. Infiltrated N. benthamiana leaves were harvested, followed by protein extraction and streptavidin beads enrichment. The enriched products were subjected to immunoblot analysis using the Streptavidin-HRP or antibodies as indicated on the bottom right of corresponding panels. Arrowheads indicate the bands of TurboID fusions. f, A Venn diagram illustrates the overlap among significantly enriched proteins identified in two sets of PL experiments depicted in (c).
