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. 2023 Aug 1;14(4):1331–1348. doi: 10.14336/AD.2023.0310

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Copyright: © 2023 Schulz et al.

this is an open access article distributed under the terms of the creative commons attribution license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 5. — Influenza virus load is enhanced in senescent human lung fibroblasts. (A) Schematic representation of infection with IAV (MOI 1) of senescent IMR-90s (IAV_TNF-α) for 8 h and 24 h. (B) qRT-PCR analysis of IL-6, IL-8, and IP-10 of quiescent mock- (MOCK_QUI) and IAV-infected (IAV_QUI), as well as uninfected senescent cells (MOCK_TNF-α) and IAV_TNF-α, was performed (N = 3). Significance was calculated with One-way ANOVA (ns p > 0.05; * p ≤ 0.05). (C) Protein levels of IL-1β, IL-6, IL-8, IFN-γ, and TNF-α were increased in IAV_TNF-α after 24 h (N = 3). Significance was calculated with One-way ANOVA (ns p > 0.05; * p ≤ 0.05). (D) Viral titer detected by standard plaque assay was increased in IAV_TNF-α compared with IAV_QUI after 24 h (N = 3). Significance was calculated with One-way ANOVA (ns p > 0.05; * p ≤ 0.05). (E) Immunofluorescence staining of IAV_TNF-α and IAV_QUI for IAV nucleoprotein, phalloidin, and DAPI showed higher signals for IAV_TNF-α. Scale bar 50 µm. (F) Transcriptomic analysis of senescent (IAV_TNF-α) and quiescent IMR-90s (IAV_QUI) after IAV infection. Overall, in the Venn diagram, the IAV_TNF-α group shares 144 RNAs with the IAV_QUI group. 113 RNAs are individually expressed in IAV_TNF-α in comparison to 195 individually expressed RNAs in the IAV_QUI. (G) Different signalling pathways are illustrated in the GO enrichment analysis of IAV_TNF-α and IAV_QUI. The IAV_TNF-α group has significant changes regarding cell motility, extracellular matrix, and signalling receptor activity. (H, I) Individually expressed genes are demonstrated in heatmaps. The genes JAK3, SOCS3, SOCS1, TNFAIP3, and CXCL6, are differentially expressed in IAV_TNF-α compared to IAV_QUI. Gene expression in heatmaps over log2 fold-change. (J) Schematic representation of the influence of an IAV infection on senescent cells and the SASP.

Figure 5. — Influenza virus load is enhanced in senescent human lung fibroblasts. (A) Schematic representation of infection with IAV (MOI 1) of senescent IMR-90s (IAV_TNF-α) for 8 h and 24 h. (B) qRT-PCR analysis of IL-6, IL-8, and IP-10 of quiescent mock- (MOCK_QUI) and IAV-infected (IAV_QUI), as well as uninfected senescent cells (MOCK_TNF-α) and IAV_TNF-α, was performed (N = 3). Significance was calculated with One-way ANOVA (ns p > 0.05; * p ≤ 0.05). (C) Protein levels of IL-1β, IL-6, IL-8, IFN-γ, and TNF-α were increased in IAV_TNF-α after 24 h (N = 3). Significance was calculated with One-way ANOVA (ns p > 0.05; * p ≤ 0.05). (D) Viral titer detected by standard plaque assay was increased in IAV_TNF-α compared with IAV_QUI after 24 h (N = 3). Significance was calculated with One-way ANOVA (ns p > 0.05; * p ≤ 0.05). (E) Immunofluorescence staining of IAV_TNF-α and IAV_QUI for IAV nucleoprotein, phalloidin, and DAPI showed higher signals for IAV_TNF-α. Scale bar 50 µm. (F) Transcriptomic analysis of senescent (IAV_TNF-α) and quiescent IMR-90s (IAV_QUI) after IAV infection. Overall, in the Venn diagram, the IAV_TNF-α group shares 144 RNAs with the IAV_QUI group. 113 RNAs are individually expressed in IAV_TNF-α in comparison to 195 individually expressed RNAs in the IAV_QUI. (G) Different signalling pathways are illustrated in the GO enrichment analysis of IAV_TNF-α and IAV_QUI. The IAV_TNF-α group has significant changes regarding cell motility, extracellular matrix, and signalling receptor activity. (H, I) Individually expressed genes are demonstrated in heatmaps. The genes JAK3, SOCS3, SOCS1, TNFAIP3, and CXCL6, are differentially expressed in IAV_TNF-α compared to IAV_QUI. Gene expression in heatmaps over log2 fold-change. (J) Schematic representation of the influence of an IAV infection on senescent cells and the SASP.