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. 2007 Mar 7;81(12):6690–6699. doi: 10.1128/JVI.02457-06

FIG. 3.

FIG. 3.

Artificial sRNAs can interfere with the silencing suppressor activity of 2b. (A) Coexpression of miR2b or shsR2b with the 2b protein reduced the activity of 2b in suppression of silencing of GFP expression in N. benthamiana 16c leaves. The GFP fluorescence images of the agroinfiltrated leaves were taken at 4 dpi under UV illumination. The bottom graph shows results of quantitative GFP fluorescence analysis averaged from two leaves for each infiltration treatment. (B) Effects of miR2b and shsR2b on the activity of 2b in enhancing the expression level of the GFP protein in WT N. benthamiana leaves. Total protein was extracted from the agroinfiltrated leaf area for each infiltration treatment at 4 dpi and subjected to Western blot analysis probed with rabbit anti-GFP antibodies. Coomassie Bright Blue staining of the large subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase indicates equal loading. (C) miR2b and shsR2b have differential effects on the activity of 2b in inhibiting systemic silencing. Images show different patterns of agroinfiltration into 16c plants and their effects on systemic silencing of GFP expression at 14 dpi. A representative plant coinfiltrated with 35S:GFP/35S:2b/35S:miR2bprec showed systemic silencing of GFP expression (frame 3) as the plant infiltrated with 35S:GFP alone (frame 1), and a plant coinfiltrated with 35S:GFP/35S:2b/U6:sh2b exhibited GFP expression in systemic leaves (frame 4), similar to that observed in the 35S:GFP/35S:2b coinfiltrated plant (frame 2).