Fig 1. Setting up of high throughput genetic screen of human ISG shRNA library to identify anti-SeV ISGs.

(A) HeLa cells stably expressing shRNA against IRF9 or a non-targeting (NT) control were pre-treated with human IFN-β for 16 h, when the cells were infected with SeV (moi:10). SeV C protein expression was analyzed by immunoblot at 16 hpi. (B) HeLa cells stably expressing shRNA against IRF9 or NT, were pre-treated with IFN-β for 16 h, when the cells were infected with SeV; 16 h later flow cytometric analyses were performed after immunostaining the cells with anti-SeV antibody. (C) A strategy to quantify percent SeV infectivity using flow cytometric procedure, as described in (B). The numbers indicate each quadrant and their respective cell population. (D) Percent SeV infectivity in NT or IRF9 shRNA-expressing HeLa, infected with SeV in the absence or the presence of IFN-β pre-treatment. (E) Our strategy to screen the human ISG shRNA library to isolate anti-SeV ISGs.