<i>NbGAPDH-A</i> does not affect the accumulations of RCNMV RNA and MP at the single-cell level.

<p>(<b>A</b>) An <i>Agrobacterium</i> culture that contained the pBICR12fsMP plasmid, that expressed movement-deficient RCNMV (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004505#ppat.1004505.s001" target="_blank">Figure S1G</a>), was diluted to OD₆₀₀ = 0.8 and infiltrated into ALSV- or ALSV/gsGAP-infected <i>N. benthamiana</i> plants (two plants for each). At 26 hpi and 43 hpi, the total RNA was extracted from the infiltrated leaves and subjected to Northern blotting using DIG-labeled riboprobes specific for the plus-strand (+) RNA1 or RNA2 of RCNMV. rRNA is an ethidium bromide-stained agarose gel image of 1 µg total RNA, which was used as the loading control. The numbers below the images represent the relative accumulation levels (means ± SE) of viral RNAs using the Image Gauge program (Fuji Film), which were calculated based on two independent experiments. Asterisk indicates a not significant (P>0.05; Student's <i>t</i>-test) difference compared with the viral RNA accumulation level in the protoplasts from ALSV-infected <i>N. benthamiana</i>. (<b>B</b>) Protoplasts prepared from ALSV- or ALSV/gsGAP-infected <i>N. benthamiana</i> plants (each two plants) were inoculated with a mixture of <i>in vitro</i> transcripts of the recombinant RCNMV RNA1, which expressed GFP from subgenomic RNA; and RNA2, which expressed MP tagged with HA (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004505#ppat.1004505.s001" target="_blank">Figure S1H</a>). Proteins extracted from 2×10⁴ protoplasts were loaded in each lane. GFP and MP-HA were detected using a rabbit polyclonal antibody against GFP and a rat polyclonal antibody against HA, respectively. RbcL is a Coomassie brilliant blue-stained gel image of proteins extracted from 2×10⁴ protoplasts, which shows the large subunit of Rubisco proteins. The numbers below the images represent the relative accumulation levels (means ± SE) of the proteins using the Image Gauge program (Fuji Film), which were calculated from two independent experiments. Double asterisk indicates a not significant (P>0.05; Student's <i>t</i>-test) difference compared with the protein accumulation level in the protoplasts from ALSV-infected <i>N. benthamiana</i>.