Figure 6

Knocking-down the expression of GRP78 by siRNA decreases the yield of infectious JE virus production. BHK-21 cells were transfected with siRNA against GRP78 or an irrelevant siRNA (negative control). The down-regulation of GRP78 was measured by Western blot analysis using antibody specific to GRP78 as shown in Figure 5. A) Transfected cell lysates were then infected with JEV at an MOI of 1. At 24 hours post-infection, the supernatants were collected to measure the amount of JE viral RNA production by RT real-time PCR as described in Material and Methods. The virus yield is expressed as a percentage of the yield obtained from cells transfected with irrelevant siRNA. B) Plaque formation by JE virus-particle collected from JEV-infected scramble siRNA treated cells or cells treated with siRNA against GRP78. C) Quantitative measurement of viral progeny produced from JEV-infected scramble siRNA treated cells or cells treated with siRNA against GRP78. The virus titer is defined as plaque-forming unit (PFU) per mL. Results are derived from three independent experiments.