Figure 4: Loss of nCBP does not interfere with PlAMV RNA accumulation at the single-cell level. | Scientific Reports

Figure 4: Loss of nCBP does not interfere with PlAMV RNA accumulation at the single-cell level.

From: Deficiency of the eIF4E isoform nCBP limits the cell-to-cell movement of a plant virus encoding triple-gene-block proteins in Arabidopsis thaliana

Figure 4

(a) Relative accumulation of PlAMV RNA at the single cell level measured by quantitative RT-PCR. Protoplasts prepared from the ncbp-1 mutant and Col-0 were transfected with a PlAMV-containing plasmid together with a GFP-expressing plasmid. Extracted total RNAs from protoplast samples at three different time points were analyzed using RT-qPCR. The level of PlAMV RNA accumulation was normalized to that of GFP mRNA in each sample. The mean level of viral RNA in Col-0 immediately after inoculation (0 hpi) was used as the standard (1.0). Error bars represent standard errors of three independent experiments. (b) Detection of viral RNAs by northern blot analysis. Protoplasts from ncbp-1 and Col-0 were inoculated with PlAMV. Total RNA (1 μg) extracted from protoplast samples at 3 dpi was subjected to northern blot analysis using specific probes. Cropped images are shown and the full-length blots are presented in Supplementary Figure S9. Uninoculated controls are indicated. Bands of ribosomal RNA are shown as loading controls. Experiments were replicated three times.

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