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. 2009 Aug 5;83(20):10460–10471. doi: 10.1128/JVI.00819-09

FIG. 8.

FIG. 8.

FIG. 8.

Localization of dsRNA and BrUTP-labeled RNA with plant and viral proteins in TuMV-infected protoplasts. N. benthamiana leaves agroinfiltrated with A. tumefaciens Agl1 containing pCambiaTunos were collected for protoplast isolation. TuMV-infected protoplasts were processed for double-label immunofluorescence with antisera raised against dsRNA and VPg-Pro (A), BrdU and VPg-Pro (B), dsRNA and RdRp (C), dsRNA and CI (helicase) (D), dsRNA and CP (E), dsRNA and eIF(iso)4E (F), dsRNA and PABP2 (G), and dsRNA and eEF1A (H). Goat anti-mouse conjugated to Alexa Fluor 568 (red, for visualization of dsRNA and BrdU) and goat anti-rabbit conjugated to Alexa Fluor 488 (green, for visualization of viral or host proteins) were used as secondary antibodies. (I) N. benthamiana leaves agroinfiltrated with A. tumefaciens Agl1 containing pCambiaTunos/6KGFP and protoplasts were processed for dsRNA immunofluorescence detection as described above. The left panels show fluorescence emitted by the green channel only, the middle panels show fluorescence emitted by the red channel only, and the right panels show the merging of the red and green channels. The inset in panel E (left panel) is a close-up view of depicted square. Scale bar, 10 μm.