ON-TARGETplus Human CASP3 (836) siRNA - SMARTpool

Protocol tips

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	Transfection was performed using DharmaFECT 4 transfection reagent (GE Dharmacon, Lafayette, CO, USA) in a 96-well white plate with a clear bottom. A smart pool of non-targeting control (NTC) siRNA (D-001206-13; GE Dharmacon) and caspase 3 siRNA (L-004307-00; GE Dharmacon) were used as negative and positive control, respectively. siRNA was diluted in DharmaFECT cell culture reagent (GE Dharmacon) and used at a final concentration of 50 nM. Transfection reagent and siRNA were mixed and incubated at 25°C for 30 minutes to form siRNA-liposome complex. Huh7 cells at 1.5 x 104 cells per well were allowed to plate onto the transfection mixture and were then incubated for 24 hours. The media were then aspirated out and the transfected cells were infected with supernatant containing DENV at MOI 10 and incubated for 48 hours. Cell viability and caspase 3 activity were measured as previously described.

Protocol tips

Transfection was performed using DharmaFECT 4 transfection reagent (GE Dharmacon, Lafayette, CO, USA) in a 96-well white plate with a clear bottom. A smart pool of non-targeting control (NTC) siRNA (D-001206-13; GE Dharmacon) and caspase 3 siRNA (L-004307-00; GE Dharmacon) were used as negative and positive control, respectively. siRNA was diluted in DharmaFECT cell culture reagent (GE Dharmacon) and used at a final concentration of 50 nM. Transfection reagent and siRNA were mixed and incubated at 25°C for 30 minutes to form siRNA-liposome complex. Huh7 cells at 1.5 x 104 cells per well were allowed to plate onto the transfection mixture and were then incubated for 24 hours. The media were then aspirated out and the transfected cells were infected with supernatant containing DENV at MOI 10 and incubated for 48 hours. Cell viability and caspase 3 activity were measured as previously described.

Publication protocol

Transfection was performed using DharmaFECT 4 transfection reagent (GE Dharmacon, Lafayette, CO, USA) in a 96-well white plate with a clear bottom. A smart pool of non-targeting control (NTC) siRNA (D-001206-13; GE Dharmacon) and caspase 3 siRNA (L-004307-00; GE Dharmacon) were used as negative and positive control, respectively. siRNA was diluted in DharmaFECT cell culture reagent (GE Dharmacon) and used at a final concentration of 50 nM. Transfection reagent and siRNA were mixed and incubated at 25°C for 30 minutes to form siRNA-liposome complex. Huh7 cells at 1.5 x 104 cells per well were allowed to plate onto the transfection mixture and were then incubated for 24 hours. The media were then aspirated out and the transfected cells were infected with supernatant containing DENV at MOI 10 and incubated for 48 hours. Cell viability and caspase 3 activity were measured as previously described.

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