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. 2024 Jul 23;14:16895. doi: 10.1038/s41598-024-66473-4

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2024

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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SARS-CoV-2 [S1S2] spike protein activates pro-Inflammatory TNFα/NF-κB signaling proteins and IL-8 expression in differentiated BCi.NS1.1 (d-BCi) epithelia. The apical surfaces of the ALI differentiated dBCi epithelia were treated with different concentrations of Wuhan-Hu-1 [S1S2] spike protein for 4 h, washed and then incubated for an additional 20 h at the Air–Liquid-Interface (ALI) condition. (a) Representative western blotting images showing the expression levels of TNFR1, TRADD, phospho-IKKα/β (Ser 176/180), IKKα + β, phospho-IκBα (Ser 32/36), IκBα, phospho-NF-κB p65 (Ser 176), NF-κB p65, and β-Actin. β-actin was used for equal loading of protein. (b–f) Quantification by densitometric analysis for the data in part (a) using BioRad Image Lab software. The data are expressed as mean ± SD (N = 5). (g) IL-8 expression in the subphase of the ALI cultures. Data are presented as means ± SD (N = 3). Statistical p values were determined with a one-way ANOVA, followed by both Holm’s and Dunnett’s post-hoc tests comparing each mean to the medium control. For the Holm test, *, p < 0.05; and ¥, p < 0.01. Dunnett’s tests were consistently significant (p < 0.05), except as noted (•).