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. 2020 Nov 12;11:581856. doi: 10.3389/fmicb.2020.581856

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Copyright © 2020 Wang, Li, Qiao, Wang, Liu, Li, Ma, Yang, Ruan, Weng, Hiscox, Stewart, Nan, Zhang and Zhou.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Recombinant PRRSV viruses bearing mutations of residues 588 and 643 of Nsp9 impaired PRRSV replication in vitro. MARC-145 Cells (A) and CRL-2843-CD163 cells (C) were infected with mutated viruses or wild type rSD16 (0.1 MOI). IFA was performed at 48 hpi with the monoclonal antibody recognized PRRSV-N protein (Clone No. 6D10) and visualized by FITC-conjugated goat anti-mouse IgG. Overlay images were visualized for cell nuclei (blue) and N protein (green). The scale bars are 100 μm. (B,D) The ratio of virus-infected cells/total cells calculated using Image J software. Asterisk (*) indicates a significant difference between rSD16 and rSD16-I588A (***p < 0.001). Pound (^#) shows a significant difference between rSD16 and rSD16-L643A (###p < 0.001).