Fig 6.

siRNA knockdown of NCL results in decreased titers of DENV. HEK293 cells were treated with NCL siRNA or NC siRNA. Twenty-four hours after siRNA treatment, the cells were infected with DENV at an MOI of 0.1, and samples were collected 72 h after infection. (A) Western blots were performed on cell lysates using antibody to NCL to verify siRNA knockdown and an antibody to actin as a loading control. (B) The titer of DENV collected from NC and NCL siRNA groups was determined in duplicate on Vero cells. Three independent experiments were used to determine significance (P < 0.01). (C) HEK293 cells were treated with NC or NCL siRNA as described for panel A. The cells were then infected with either EMCV (MOI, 0.001) or VSV (MOI, 0.001) for 24 h. The titers of samples were determined in quadruplicate on Vero cells using the limiting-dilution method, and the 50% tissue culture infectious dose (TCID₅₀) was calculated. Two independent experiments were used to determine significance (P < 0.01). The error bars indicate SD.