Fig 9.

NCL does not significantly affect release of DENV particles. (A) Western blot of HEK293 cells infected with DENV (MOI, 3), followed by treatment with AS1411 or the NC. Cell supernatants were collected at time zero and at 24-h intervals until 96 h postinfection. Virus was purified through a sucrose cushion, and samples were analyzed by SDS-PAGE, followed by Western blotting of DENV C and DENV E proteins. The data are representative of three independent experiments. (B) qRT-PCR of viral RNA extracted from cell supernatants of DENV-infected HEK293 cells (as described for panel A) and no-virus control (NVC). Samples were analyzed using primers to DENV and normalized to norovirus G2 RNA added to the sample prior to extraction. The data are representative of two independent experiments performed in triplicate. (C) Western blot of HEK293 cells treated with siRNA to NCL or the NC, followed by infection with DENV (MOI, 3). Cell supernatants were collected at 72 h postinfection. Virus was purified through a sucrose cushion, and samples were analyzed by SDS-PAGE, followed by Western blotting of DENV C and DENV E proteins. (D) qRT-PCR of viral RNA extracted from cell supernatants of DENV-infected HEK293 cells after treatment with siRNA for NCL or negative-control siRNA. Samples were analyzed using primers to DENV and normalized to norovirus G2 RNA added to the sample prior to extraction. The data are representative of two independent experiments performed in triplicate. The error bars indicate SD.