FIG. 2.

(Top). Localization of Br-RNA and dsRNA in typical PCV-infected BY-2 tobacco protoplasts. PCV-infected (A, B, and C) or mock-inoculated (D and E) protoplasts were treated with actinomycin D added at 17 hpi and then incubated for 1 h prior to addition of BrUTP and further incubated for 6 h. The protoplasts were then immunolabeled with mouse BrUTP-specific antibodies and dsRNA guinea pig antibodies, which were then detected with, respectively, Alexa 488 (A and D)- and Alexa 568 (B and E)-labeled secondary antibodies. BrUTP labeling is in green, and dsRNA labeling is in red. (C) Superimposition of the images to the left. The confocal images were collected with a focal depth of 0.45 μm. Bar, 10 μm.