Figure 4

RT-PCR analysis of host gene expression and Western blotting analysis of BaMV coat protein accumulation in ACAG1- and ACAG8-knockdown plants. ACAG1- and ACAG8-knockdown plants were inoculated with viral RNA. The GFP plant was included as the negative control. RNA and protein extracts of leaves inoculated with viral RNA were harvested on 5 dpi. The RNA extracts were subjected to ACAG1- (A) or ACAG8-specific (C) semi-quantitative RT-PCR. RT-PCR data was normalized to the levels of actin. Protein extracts were analyzed for BaMV coat protein accumulation by Western blotting (B and D). The Rubisco large subunit (rbcL) was included as the loading control for normalization. For all experiments, the levels detected in the GFP control plants on 5 dpi were set as 100%. Representative results are shown under the statistical results showing the average of the relative levels of AGAC1 mRNA (A), ACAG8 mRNA (C) and, BaMV coat protein (B and D) with standard deviations derived from at least three independent experiments. Asterisks indicate statistically significant differences compared with the control indicated as GFP (**p < 0.05, ***p < 0.001).