Figure 5

Virus and GFP accumulation in wild type and 10.6.1 transgenic N. benthamiana plants inoculated with different viruses. Extracts of inoculated leaves (inoc.), the leaf immediately above the inoculated ones (+ 1 leaf), and the fourth leaf above the inoculated ones ( + 4 leaf) from 10.6.1 transgenic or wild type N. benthamiana plants inoculated with PPV (panel A), TVMV (panel B) or CMV (panel C) displaying (S) or not displaying (NS) symptoms (either single plants or pools from two or three plants), collected at 18 days post inoculation, were subjected to western analysis with the indicated antibodies (TVMV antibody also reacts with PPV CP). One mock inoculated 10.6.1 plant that developed an spontaneous infection (S) and a pool of three of these plants that did not show symptoms of amplicon-derived infection (NS) were also analyzed (panel A). Samples from a 10.1.7 transgenic plant, expressing the PPV-NK-GFP amplicon from germination, and from a healthy N. benthamiana plant (wt Nb) were used as positive and negative controls, respectively. The blots stained with Ponceau red showing the rubisco protein are presented as loading controls.