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. 2023 Sep 27;14:6030. doi: 10.1038/s41467-023-41442-z

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — A Arrayed siRNA screen approach in A549 cells reverse-transfected in n = 2 biologically independent samples with gene-targeting, non-targeting (NT) or IAV NP-targeting siRNA, and infected with Influenza A/WSN/1933 H1N1 (MOI 0.1, 24 hours). Cell viability (live-cell staining) and percent IAV infection (%NP-positive (%NP + ) cells; immunostaining for IAV NP) were quantified by flow cytometry. Correlation plots comparing: B the cell viability against IAV infection for each siRNA from each biological duplicate; or C the variation in IAV infection between the biological duplicates. Log2FC was calculated by normalizing %viable or %NP+ cells for each siRNA against the mean of multiple replicate-matched NT siRNA (siRNA/mean NT) (green dots=IAV NP-targeting siRNA, black dots=NT siRNA, grey dots=experimental gene-targeting siRNA). Distribution of log2FC in IAV infection for (D) 212 PPI targets and (E) 78 PH targets, plotted as the mean of n = 2 biologically independent samples per target. The log2FC in IAV infection was calculated for each siRNA against the mean replicate-matched NT siRNA (blue dots=pro-viral factors (mean log2FC < -2), red dots=antiviral factors (mean log2FC > 2), grey dots=no/weak phenotype; green dot=IAV NP-targeting siRNA; black dot=NT siRNA; error bars represent standard deviation). F Distribution of log2FC in SARS-CoV-2 infection for 44 IAV PPI targets plotted as the median of six replicates (n = 2 biologically independent samples, each in n = 3 technical replicates) per target. The log2FC in SARS-CoV-2 infection was calculated for each siRNA against a replicate-matched NT siRNA (blue dots=pro-viral factors (median log2FC < -2), red dots=antiviral factors (median log2FC > 2), grey dots=no/weak phenotype; green dot=ACE2-targeting siRNA; black dot=NT siRNA; error bars represent median absolute deviations (MAD)). G Bar chart of pro-viral and antiviral factors for IAV and SARS-CoV-2 screens plotted as the mean log2FC in IAV infection (data re-plotted from D and E; error bars represent standard deviation) and the median log2FC in SARS-CoV-2 infection (data re-plotted from F; error bars represent MAD).