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. 2024 Feb 16;221(3):e20231144. doi: 10.1084/jem.20231144

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© 2024 Hassert et al.

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Figure S1. — Gating strategies for T_RM, T_CM, and T_EM. 3 min prior to euthanasia, mice were administered 2 μg of BV421 labeled anti-CD45.2 to discriminate vascular from tissue cells. (A) For lung T_RM, cells were gated on lymphocytes, live, and IV-negative cells that express CD8, followed by CD90.1 to identify P14 cells. Lung T_RM are identified as being double positive for CD69 and CD103. (B) Splenocytes were gated on lymphocytes, single cells that are CD8⁺. CD11a is used to identify antigen-experienced cells, which are then further subset into P14 cells using CD90.1. T_CM and T_EM are differentiated using CD62L and CD27. (C–F) Flow cytometric analysis was utilized on whole spleens to enumerate the total number of (C) CD8⁺ T cells, (D) P14 cells, (E) T_CM P14 cells, and (F) T_EM P14 cells. ANOVA was utilized to determine statistically significant differences between groups. Data are pooled from two independent experiments (n = 6–7).