Interplay of type I and II interferons promotes T _SCM cell differentiation via multiple mechanistic pathways. (A–C) P14 cell positioning at d8 of acute LCMV infection in wild-type, Ifnar^−/−, Ifng^−/−, or 2xIFN^−/− mice. Images and graphs are representative of three individual experiments with three to five mice in each group per experiment. (A) LSFM micrographs of intact lymph nodes. Images are 200-µm longitudinal slices through the lymph node center. Scale bars represent 200 µm. Images show GFP-P14 cells (yellow), B220 (B cells; cyan), and CD31 (vessels; magenta). (B) Graph summarizing density of P14 cells within the 3D lymph node, from periphery (EVF = 0) to center (EVF = 1). The colored dashed line indicates multiple t tests between P14 cell density in the three experimental conditions against the wild type for each EVF value. The black dashed line indicates P = 0.05. Data are the mean ± SEM. (C) Graphs summarizing density of P14 cells within indicated regions, IFRs (EVF 0.1–0.3) and T cell paracortex (EVF 0.7–0.9). Data are the mean ± SEM. Statistical differences were analyzed using an unpaired t test. (D) Graphs summarizing T_EFF (KLRG1⁺SLAMF6⁻) and T_SCM (KLRG1⁻SLAMF6⁺) cell populations in each of the four conditions. (E) Graph summarizing the PFU from viable virus in spleens of mice in each genotype. The dashed line indicates viral plaque LOD. Each dot in D and E represents a single mouse. Data are the mean ± SEM. Statistical differences were analyzed using one-way ANOVA tests. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Fig. S4 shows additional supporting data. LOD, limit of detection.