Extended Data Fig. 2: Vacuolar localization of pathogen upregulated proteins as putative VSR cargo proteins.

(a) Transcript levels (log₂) of genes, function as putative VSR cargo proteins, in WT plants after inoculation of the Pto DC3000 avrRpm1 or Pto DC3000 at indicated time of post inoculation. Heatmap displays the relative expression (log₂FC) of the genes in WT plants after inoculation with Pto DC3000 avrRpm1 or Pto DC3000 at different times points after inoculation relative to their expression before inoculation (0 h). (b) Subcellular localization of secYFP and Aleurain-GFP as controls. secYFP showed ER pattern, without any fluorescence signal in the vacuole. Vacuolar localized protein Aleurain-GFP colocalized with the PVC/MVB marker mRFP-VSR2 in punctae and showed diffused fluorescence signals in vacuole. Separated images of each channel in the white outline area are shown on the right side (from top to bottom: GFP/YFP, mRFP, and merged). Scale bars, 10 μm. (c) Colocalization of GFP/YFP-tagged putative VSR cargo in Papain-Like Cysteine Proteases family with the PVC/MVB marker mRFP-VSR2 was analyzed with a confocal microscope in protoplasts of Arabidopsis suspension cells. Separated images of each channel in the white outline area are shown on the right side (from top to bottom: GFP/YFP, mRFP, and merged). Scale bars, 10 μm. (d) Subcellular localization analysis of two Pathogenesis-related proteins (PR3 and PR4), three peroxidases (PRX33, PRX34, and PRX37), CPY, PEP4, and GRX. Colocalization of GFP/YFP-tagged proteins with the PVC/MVB marker mRFP-VSR2 was analyzed with a confocal microscope in protoplasts of Arabidopsis suspension cells. Separated images of each channel in the white outline area are shown on the right side (from top to bottom: GFP/YFP, mRFP, and merged). Scale bars, 10 μm. Similar confocal imaging results to those in (b–d) were obtained from three independent experiments.