Effects of siRNA silencing of host chaperones on RVFV levels.

<p>(A) Silencer Select siRNA library (Life Technologies) was used to serially assess the importance of identified chaperones in RVFV infection. HeLa cells were transfected with siRNA molecules targeting chaperone proteins, negative control targets (GAPDH, scrambled siRNA) or “no siRNA” controls. Following 48 hr incubation, cells were infected with the MP-12 strain of RVFV and the extent of viral infection assessed by high content imaging, as described in the Methods section. The down-regulation of chaperone proteins indicated in the figure resulted in statistically significant effect on viral infection when compared to the scrambled siRNA, as indicated by the asterisk symbols (P<0.05). Negative controls corresponding to GAPDH, scrambled siRNA and no siRNA control did not impact RVFV infection. The experiment was repeated in three independent trials with samples assayed in duplicate. The displayed data is a representative of the entire sample set. (B) The effects of siRNA treatments on RVFV titers were also measured using qRT-PCR analysis. For each target HSP with a duplicate siRNA set, the siRNA construct with the highest knockdown efficiency was analyzed. HeLa cells were transfected with the siRNA constructs and after 48 hours the transfected cells were infected with the MP-12 strain. The culture supernatants were analyzed 24 hours after infection. Asterisks designate significant changes in viral levels (P<0.05).