Fig. 7.
Effect of NALP3, ASC and caspase-1 siRNA on IL-1β secretion. (a–c) Mock-infected and HCV-infected cells were transfected with siRNA against GFP, NALP3, ASC and caspase-1 as described in Methods. At day 4, total cellular RNA was extracted using TRIzol, cDNA was synthesized and QRT-PCR was performed using GFP-, NALP3-, ASC- and caspase-1-specific primers (Supplementary Table S1). 18S rRNA was used as an internal control. The data shown here represent the means+sd of at least three independent experiments performed in triplicate. *, P<0.05 compared with GFP siRNA-transfected control cells. (d) Effect of NALP3, ASC and caspase-1 siRNA on IL-1β secretion. Cell culture supernatants from siRNA-transfected mock-infected and HCV-infected cells were subjected to IL-1β ELISA. *, P<0.05 compared with mock-infected control cells. **, P<0.05 compared with GFP siRNA-transfected cells. (e) Cytotoxicity assay. Equal volume of cell culture medium from mock-infected and HCV-infected cells, transfected with GFP, caspase-1 and NALP3 siRNA were subjected to cytotoxicity assay using CytoTox-ONE. RFU, Relative fluorescence unit.