FIG. 2.
Transient transfection of amiRNAs and their effect on cell viability. (A) Cells seeded in a 96-well plate were infected with JEV at a MOI 5. Three hours postinfection, the cells were transfected with three different concentrations of amiRNAs (100, 500, and 1,000 ng) of single amiRNA per well. After 48 hpi, MTT reagent was added, and absorbance was measured at 570 nm. Results represent three independent experiments. (B) Cells were seeded in a 6-well plate and were transfected with four different concentrations of amiRNAs (50, 250, 500, and 1,000 ng) of single amiRNA per well. After 24 h, amiRNAs expression was monitored by checking eGFP expression under a fluorescence microscope. Representative images of amiRNA-treated HEK293T cells at 10 × magnification are shown. (C) RT-PCR analysis of four ISG (Ifitm1, Oasl1, Oasl2, and Isg15) mRNA expression. Gapdh gene expression was used to normalize the ISG expression. emGFP, emerald green fluorescent protein; hpi, hours postinfection; IFN, interferon; ISGs, IFN-stimulated genes; MOI, multiplicity of infection; qRT-PCR, quantitative real time polymerase chain reaction. Color images available online at www.liebertpub.com/nat