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NS1 variants and their characterization.

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posted on 2015-05-15, 03:44 authored by Woo-Chang Cheong, Hye-Ri Kang, Hyunyee Yoon, Suk-Jo Kang, Jenny P.-Y. Ting, Moon Jung Song

Plasmids expressing the NS1 variants NS1-HK and NS1-WSN were transiently transfected into HEK293T cells (A and B). (A) Expression of NS1 variants in the cells was examined by western blot analysis using an anti-MYC antibody. Tubulin was used as a loading control. (B) HEK293T cells expressing NS1 variants were fixed at 24 hr post-transfection, immunostained with anti-MYC (red) for NS1 detection, and examined under a confocal laser scanning microscope. Nuclei were stained with DAPI (blue). Scale bar, 20 μm. (C) Transfection of HEK293T cells was performed in triplicate by using the reporter construct (IFN-β-luc) and the NS1 expression plasmids (300 and 500 ng); the β-galactosidase expression plasmid served as an internal control. At 24 hr post-transfection, 20 HA of SeV was infected. After 16 hr infection, IFN-β-luc reporter activities were measured and normalized to β-galactosidase activities. Statistical analysis was performed using Student’s t-test (*** denotes a p-value of <0.005.).