Fig. 4

Accumulation of GFP mRNA and suppressor proteins in infiltrated N. benthamiana 16c leaves at 3 days after infiltration (dpi). a GFP levels in tissues co-infiltrated with cotton leafroll dwarf virus P0 proteins (P0^CL-ARG, P0^CL-PV1, P0^CL-Acr9, P0^CL-Hol1, P0^CL-Ima2, P0^CL-Ipa4, P0^CL-Pal3 or P0^CL-Pm1), potato leafroll virus P0 (P0^PL-AU), empty vector or mock-inoculated were detected with real-time PCR. Error bars indicate standard deviation of GFP mRNA in three biological repeats. Normalized value obtained in the mock sample was arbitrary set to 1 and all the other values compared to it. Data was normalized with Ubi3 and EF-1 reference genes. Asterisks indicate values that are statistically different from the control mock construct, with p-values varying from 0.0002 to 0.0478. b Western blot showing the accumulation of suppressor proteins. Protein extracts from all the three biological replicates from each construct used for real-time PCR were run in SDS-PAGE, transferred to membranes and probed with Myc-tag specific antibodies. Gel loading was observed by Ponceau staining. Non-infiltrated plants (NI) were used as negative controls. Top left panel: accumulation of the control proteins P0^PL-AU and tombusvirus P19 and the negative control (infiltrated with the empty vector). Top right panel: accumulation of P0^CL-ARG, P0^CL-PV1 and P0^CL-Acr9 proteins. Bottom left panel: accumulation of P0^CL-Hol1, P0^CL-Ima2 and P0^CL-Ipa4 proteins. Bottom right: accumulation of P0^CL-Pal3 and P0^CL-Pm1 proteins