Analysis of RNA silencing suppression function of individually deleted or point-mutated NSs proteins by agroinfiltration in <i>N</i>. <i>benthamiana</i> plants.

<p>The leaf areas agroinfiltrated with each deleted <b>(A, upper panel)</b> or point-mutated <b>(B, upper panel)</b> NSs constructs were examined under UV and white light. The GFP fluorescence was recorded at 4 days after co-infiltration (dpa) of <i>Agrobacterium</i> separately carrying pBA-GFP (the expresser), pBA-GFi (the silencing inducer) and individual constructs with deleted or point mutated NSs proteins described in Fig <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0126161#pone.0126161.g001" target="_blank">1A</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0126161#pone.0126161.g001" target="_blank">1B</a>. Western blotting was performed for the detection of individually deleted <b>(A, lower panel)</b> or point-mutated <b>(B, lower panel)</b> NSs proteins expressed at 4 dpa. NSs monoclonal (MAb) and NSs polyclonal antibodies (PAb) were used for detecting NSs protein or NSs protein in which the common epitope was mutated, respectively. Expression levels of point-mutated NSs proteins, NSs mRNA, GFP and GFP mRNA were detected at 4 dpa <b>(C)</b>. Coomassie blue-stained RuBisCO protein was used as loading controls for proteins and 18S rRNA used as loading controls for RNAs.