EPS8 facilitates uncoating of influenza A virus

Gloria P. Larson; Vy Tran; Shuiqìng Yú; Yíngyún Caì; Christina A. Higgins; Danielle M. Smith; Steven F. Baker; Sheli R. Radoshitzky; Jens H. Kuhn; Andrew Mehle

FVG-R infections were performed by inoculating A549 cells with virus diluted in Opti-MEM I (Invitrogen, 31985070) supplemented with 0.2% FBS. Viral gene expression was measured 8 hpi using a Renilla luciferase assay system (Promega). Infections with JUNV (Romero), EBOV, and MARV (Ci67) and infections with RVFV (ZH501) and VEEV (IC-SH3) were conducted under Biosafety Laboratory 4 and 3 conditions, respectively. Cells in 96-well format (30,000 cells per well) were infected at the indicated MOIs. After 1 hour, the inocula were removed, cells were washed with PBS, and replenished with fresh growth media. VEEV and RVFV-infected plates were fixed in formalin 20 hours post-inoculation. All other infected plates were fixed 48 hours post-inoculation. Antigen staining and high-content quantitative image-based analysis were performed as previously described (Radoshitzky et al., 2010 (Radoshitzky et al., , 2016 . The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/592485 doi: bioRxiv preprint EPS8 facilitates uncoating of influenza A virus 11 Larson, Tran, et al.