Fig 4. HCPro^WD is defective in recruiting VCS to PGs.

(A-F) Localization of Strep-RFP-tagged -HCPro, YFP -tagged-VCS and CFP tagged -P0 in N. benthamiana epidermal cells. Agrobacterium carrying either HCPro^WT-Strep-RFP or HCPro^WD-Strep-RFP was infiltrated at OD₆₀₀ = 0.1, while Agrobacteria harboring VCS-A^YFP, VCS-B^YFP and VCS-C^YFP were each infiltrated at OD₆₀₀ = 0.1 (together denoted as VCS^YFP). Agrobacterium harboring P0^CFP, infiltrated at OD₆₀₀ = 0.1, was expressed as another marker for PGs. Samples were analyzed at 3 dpi by confocal laser scanning microscopy in sequential scanning mode. (A) P0^CFP, (B) VCS^YFP, (C) HCPro^WT-Strep-RFP, (D) P0^CFP, (E) VCS-^YFP and (F) HCPro^WD-Strep-RFP. (G-L) Localization of HCPro^{WT/WD-Strep-RFP} and VCS^YFP and their overlays in N. benthamiana epidermal cells. Experimental details are otherwise the same as in Fig 4A–4F, except for the absence of P0^CFP. (G) HCPro^WT-Strep-RFP, (H) VCS-^YFP and (I) Overlay of HCPro^WT-Strep-RFP and VCS^YFP. (J) HCPro^WD-Strep-RFP, (K) VCS^YFP and (L) Overlay of HCPro^WD-Strep-RFP and VCS^YFP. Degree of co-localization between HCPro^WT-Strep-RFP / HCPro^WD-Strep-RFP and VCS^YFP is calculated from representative images and presented in S6 Fig. (M-R) Epifluorescence microscopy showing the distribution of HCPro^{WT/WD-Strep-RFP} and P0^YFP within a representative area of examination. HCPro and P0^YFP, which both typically co-localize in PGs, were co-infiltrated to visualize the PGs. The experiment was conducted similarly as Fig 3E and 3F. The only exception was that HCPro^{WT/WD-Strep-RFP} was used instead of HCPro^WT/WD. Images were acquired at 3 dpi with a 10X objective. HCPro^{WT/WD-Strep-RFP} was visualized under an RFP filter (M, P respectively). P0^YFP from the corresponding sample sets was visualized under the FITC filter (N, Q respectively), and their overlays are presented in (O, R), respectively.