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Figure 6 | Virology Journal

Figure 6

From: Actively replicating West Nile virus is resistant to cytoplasmic delivery of siRNA

Figure 6

WNV RNAi resistance is dependent of viral translation early in infection. A. Puromycin does not interfere with RNAi. Huh7.5 cells were mock-treated or treated with 6 μg/ml puromycin and transfected with Cap or Cap Mut siRNA for 4 hours. Cells were washed twice, the puromycin replaced, and cells were infected with WNV at MOI 0.1. Four hours later cells were washed twice and replaced with medium that lacked puromycin. Forty-eight hours after infection, cells were collected and analyzed for WNV envelope protein expression by flow cytometry. Fold inhibition was calculated as described above. The results are an average of three independent experiments and error bars indicate standard error of the mean. B. Puromycin time course. Huh7.5 cells were infected with WNV (MOI = 0.01), and puromycin was added at the indicated times before or after infection. At 9 hours post-infection cells were washed and transfected with Cap or Cap Mut siRNA. Forty-eight hours later, WNV envelope protein expression was assessed by flow cytometry. Fold inhibition was calculated as described in Fig 3B. The results are an average of three independent experiments and error bars indicate standard error of the mean.

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