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. 2023 Jul 19;14:4331. doi: 10.1038/s41467-023-40107-1

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Control, CD2rtTA x CA-IKK2 or CD2rtTA x DN-IKK2 mice were infected with influenza X31(1000 pfu). From 5–30 days post-infection (d.p.i.), mice were fed a DOX or a control diet. Numbers (#) in mediastinal lymph nodes at 30 d.p.i of (b) NP_366–374-specific CD44^hi, CD62L^hi T_MEM cells (CD8⁺, Db-NP-tet⁺, CD44⁺, CD62L⁺), n = 9 (control), n = 5 (DOX control), n = 6 (CA-IKK2^ON), n = 8 (DN-IKK2^ON) mice pooled from 3 independent experiments (c) NP_366–374-specific CD44^hi, CD62L^lo (CD8⁺ Db-NP-tet⁺ CD44^hiCD62L^lo) T_MEM cells. n = 6 (control), n = 6 (DOX control), n = 5 (CA-IKK2^ON), n = 6 (DN-IKK2^ON) mice pooled from 4 independent experiments. (d–g) Lung T_RM were identified in X31-infected mice by intravascular (IV) staining with CD45.2 to discriminate vascular (IV. CD45.2⁺) and parenchyma resident (IV CD45.2⁻). The frequency (%) and numbers of influenza (d) NP_366–374 specific CD8⁺ T_RM. For %, #: n = 6, 10 (control), n = 6, 7 (DOX control), n = 5, 7 (CA-IKK2^ON), n = 7, 8 (DN-IKK2^ON) mice pooled from 2 independent experiments. e PA_224–233 specific CD8⁺ T_RM. n = 11, 8 (control), n = 12, 4 (DOX control), n = 6, 9 (CA-IKK2^ON), n = 7, 5 (DN-IKK2^ON) mice pooled from 2 independent experiments. f Representative dot plots showing frequencies and numbers of cells with the CD69⁺, CD103⁺ phenotype among NP_366–374 specific CD8⁺ T cells (top graphs) or among IV negative (bottom graphs). For %, #: n = 10, 10 (control), n = 5, 3 (dox control), n = 10, 8 (CA-IKK2^ON), n = 5, 5 (DN-IKK2^ON) mice pooled from n = 2 independent experiments for top graphs. n = 5, 3 (control), n = 3, 3 (DOX control), n = 4, 3 (CA-IKK2^ON), n = 4, 3 (DN-IKK2^ON) mice pooled from 2 independent experiments for bottom graphs. g Control or CD2rtTA x CA-IKK2 (CA-IKK2^ON) mice were infected with 2 × 10⁶ pfu VSV, fed a DOX-containing diet from 5 to 30 d.p.i. At 30 d.p.i. VSV-specific CD8⁺ T_RM (Kb-N-tet⁺ CD8⁺ CD45.2⁻) were identified in the lungs and kidneys of infected mice by IV staining. n = 5 (control), n = 6 (CA-IKK2^ON) mice pooled from 2 independent experiments. Bars represent mean values ± SD. P-values were determined by a two-tailed unpaired t test, n.s. not significant. Source data provided as a source data file.