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. 2016 Apr 29;90(10):5119–5131. doi: 10.1128/JVI.00024-16

FIG 6.

FIG 6

PPV 6K1 punctate inclusions are associated with chloroplasts and colocalize with VRC elements 6K2 and NIb during viral early infection. (A) Time course of 6K1 punctate inclusions (36, 48, and 72 hpi [panels I to III, respectively]) in association with chloroplasts in N. benthamiana leaves infiltrated with WT::6K1GFP. Green and red fluorescence signals represent 6K1-GFP and chlorophyll autofluorescence, respectively. Arrowheads, 6K1-GFP punctate inclusions. Bars, 10 μm. (B) 6K1 punctate inclusions colocalize with 6K2 in association with chloroplasts in N. benthamiana leaves infiltrated with pVPH::6K1GFP (which was constructed by removal of the mCherry expression cassette of WT::6K1GFP) and further infiltrated with pGWB451-6K2. Green fluorescence and red fluorescence represent 6K1-GFP and 6K2-RFP, respectively. Chl, chlorophyll autofluorescence (blue). Asterisk, the nucleus. Bars, 5 μm. (C) 6K1 punctate inclusions colocalize with the NIb protein (viral RNA-dependent RNA polymerase) in association with chloroplasts in N. benthamiana leaves infiltrated with pVPH::6K1GFP and further infiltrated with pCaMterX-NIb-mCherry. Green fluorescence and red fluorescence represent 6K1-GFP and NIb-mCherry, respectively. Chl, chlorophyll autofluorescence (blue). Arrowheads and arrows, 6K1-GFP inclusions and NIb punctate structures, respectively. Bars, 10 μm.