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. 2021 Sep 3;13(9):1760. doi: 10.3390/v13091760

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Induction of innate immune responses in vivo by recombinant viruses encoding H5N1 NS1 and/or PA variants: Groups of 7-to-8-week-old A129 male mice (N = 8/group) were infected with 1000 (10³; A) or 10,000 (10⁴; B) FFU/mouse of the indicated viruses (PA_OR/NS1_OR, PA_OR/NS1_CIR, PA_CIR/NS1_OR, and PA_CIR/NS1_CIR) or left mock-infected (N = 4). Mice were sacrificed at 2 (N = 4) and 4 (N = 4) d p.i. Left lungs were collected and total RNA was extracted to quantify the levels of TNF (left panels), CCL2 (middle panels), and IFIT2 (right panels) by RT-qPCR. Levels of expression in infected mice were compared to those in mock-infected mice. *, p < 0.05; **, p < 0.005; ***, p < 0.0005; ****, p < 0.0001 (PA_OR/NS1_OR vs. PA_OR/NS1_CIR, PA_CIR/NS1_OR, and PA_CIR/NS1_CIR) using 2 way ANOVA (n = 4 per time point). Non-significant when p > 0.05.