Optimization of siRNA concentration.

<p>A) Vero E6 cells were transfected with Chik-1 and Chik-5 siRNAs individually as well as in combination at different concentration (10, 50, 100, 150 and 200 pmol). Total RNA was isolated 24 h after virus infection. Amount of CHIKV RNA was detected by measuring E3 RNA copies by real-time RT-PCR. siRNA Chik-1 and Chik-5 treatment at a concentration of 10 pM failed to reduce CHIKV RNA copies. Whereas all other concentrations of Chik-1 and Chik-5 significantly reduced the viral RNA copies. Values are given as mean log₁₀ RNA copies/well ± SD. Significance ANOVA, Dunnett's test: *<i>p</i><0.05; **<i>p</i><0.01. <b>B</b>) Effect of Chik-1 and Chik-5 siRNAs on DENV-2 growth: Vero-E6 cells were infected with DENV-2 and Chik-1 (100 pmol) CHIK-5 (100 pmol) and Comb-siRNA (100 pmol) transfected after 2 h p.i. and DENV-2 titers were measured by plaque assay. <b>C</b>) Effect of Chik-1 and Chik-5 siRNAs on Chandipura virus (CHPV) growth: Vero-E6 cells were infected with CHPV and Chik-1 (100 pmol) CHIK-5 (100 pmol) and Comb-siRNA (100 pmol) transfected after 2 h p.i. and CHPV titers were measured by plaque assay <b>D</b>) Effect of siRNA–Hiperfect complex treatment on survival of Vero-E6 cells was accessed by MTT assay. siRNAs were used in 100 pmol concentration.