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

Figure 3.

Figure 3.

TNF-α production by macrophages during infection leads to cellular senescence in human lung fibroblasts. (A) Infection of the hMDM was performed with IAV (H1N1, PR8) for 8 h and 24 h and MOI 5. After 24 h of infection, hMDM produced high levels of the cytokines and chemokines TNF-α, IFN-γ, IP-10, IL-1β, and IL-6 (N = 4). Significance was calculated with the Mann-Whitney U test (* p ≤ 0.05). (B) Immunofluorescence staining of mock- and IAV-infected hMDM of IAV-specific nucleoprotein, CD68, and DAPI. Scale bar 50 µm. (C) Schematic representation of treatment of the IMR-90s with supernatants of IAV-infected hMDM (conditioned medium, CM). (D) Using SA β-gal staining, we detected a positive signal in the IMR-90s treated with conditioned media (SENCM). Scale bar 200 µm. (E) After CM treatment (day 8), qRT-PCR analysis was performed, and CDKN1A, and MMP3 were significantly increased. Significance was calculated with the Mann-Whitney U test (ns p > 0.05; * p ≤ 0.05). (F) Protein levels were determined, and the pro-inflammatory factors IL-1β, IL-6, IL-8, IL-18, and IFN-γ were significantly upregulated (N = 4). Significance was calculated with the Mann-Whitney U test (* p ≤ 0.05).