Fig. 8

P0 suppresses GFP-induced RNA silencing. a 16c N. benthamiana plants co-infiltrated with A. tumefaciens cultures expressing GFP (35S-GFP) and vector control (Vec), BYCV P0 (P0), or TBSV 19 (P19, a positive control) were photographed under UV light at 4 dpi. Bars = 2 cm. b qRT-PCR analysis of relative GFP expression levels in the agroinfiltrated leaf patches from a. Three plants were analyzed for each treatment; this experiment was repeated three times with similar results. Error bars represent ± SD (n = 3), and NbActin2 was used as an internal reference. c western blotting analyses of the GFP protein accumulation in the agroinfiltrated leaf patches as indicated in (a). d The 16c N. benthamiana plants co-infiltrated 35S-GFP and PVX, or PVX-P0, or PVX-HC-Pro, were photographed under UV light at 7 dpi. Bars = 2 cm. e qRT-PCR analysis of relative GFP expression levels in the agroinfiltrated leaf patches from (d). Error bars represent ± SD (n = 3), and NbActin2 was used as an internal reference. Student’s t-test was used to analyze each data group statistically, and double asterisks indicate significant statistical differences (**P < 0.01) between the two treatments. f western blotting analyses of GFP and PVX CP accumulation in the agroinfiltrated leaf patches as indicated from (d). g The 16c N. benthamiana plants co-infiltrated 35S-GFP and PVX, or PVX- or PVX-HC-Pro, were photographed under UV and white light at 20 dpi. Bars = 4 cm. h qRT-PCR analysis of relative GFP expression levels in systemic leaves from (g). Error bars represent ± SD (n = 3), and NbActin2 was used as an internal reference. Student’s t-test was used to analyze each data group statistically, and double asterisks indicate significant statistical differences (**P < 0.01) between the two treatments. i western blotting analyses of GFP and PVX CP accumulation in systemic leaves from (g). Western blotting analyses of the Actin accumulation with a specific anti-Actin antibody served as a loading control (c), (f), and (i). At least 6 plants were used in each independent systemic silencing experiment. The agroinfiltration experiment. qRT-PCR and Western blotting analyses were repeated three times with similar results in this figure