Figure S3.
DC gating strategy and increased CXCR3 ligand expression and TSCMcell differentiation during live and heat-inactivated acute LCMV infection following IFNAR blocking. Related to Fig. 5. (A) Gating strategy to identify cDC1, cDC2, and moDC cell populations. Colored boxes indicate the final gate and REX3 expression for each subset. (B) Frequency and total cell number of Cxcl9-RFP+ and Cxcl10-BFP+ DC subsets at d4 and d8 after LCMV Armstrong infection. Data are representative of three individual experiments with three to four mice in each group. Data are the mean ± SEM. Each dot represents a single sample. Statistical differences were analyzed using unpaired t tests. (C) Images from Fig. 5 H. LSFM micrographs of intact REX3 lymph nodes at d8 of infection in control and d0–1 IFNAR-blocked conditions. Images are 200-µm longitudinal slices through the lymph node center. Scale bars represent 100 µm. The dashed line indicates the lymph node outline. Individual REX3 reporters and merge images show Cxcl9-RFP (magenta) and Cxc10-BFP (cyan). Images are representative of three individual experiments with at least four mice in each group in each experiment. (D and E) Analysis of Cxcl9-RFP and Cxcl10-BFP expression in DC subsets within peripheral lymph nodes of mice at d8 following challenge with either live or heat-inactivated (inactive) LCMV Armstrong. Half of each group received d0–1 treatments of IFNAR block. Data are representative of three individual experiments with three mice per group per experiment. Data are the mean ± SEM. (D) Graphs summarizing the graded frequencies of Cxcl9-RFP+ and Cxcl10-BFP+ DC subsets at d8 following acute LCMV Armstrong infection. (E) Frequency and total cell numbers and gMFI of chemokine reporter expression in each immune challenge condition. Each dot represents a single mouse. Data are the mean ± SEM. Statistical differences were analyzed using unpaired t tests. (F) Graphs summarizing the mean frequencies ± SEM of TEFF and TSCM cells in each condition. Data are representative of three independent experiments with three to four mice in each group. Statistical differences were analyzed using unpaired t tests. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

DC gating strategy and increased CXCR3 ligand expression and T SCM cell differentiation during live and heat-inactivated acute LCMV infection following IFNAR blocking. Related to Fig. 5. (A) Gating strategy to identify cDC1, cDC2, and moDC cell populations. Colored boxes indicate the final gate and REX3 expression for each subset. (B) Frequency and total cell number of Cxcl9-RFP+ and Cxcl10-BFP+ DC subsets at d4 and d8 after LCMV Armstrong infection. Data are representative of three individual experiments with three to four mice in each group. Data are the mean ± SEM. Each dot represents a single sample. Statistical differences were analyzed using unpaired t tests. (C) Images from Fig. 5 H. LSFM micrographs of intact REX3 lymph nodes at d8 of infection in control and d0–1 IFNAR-blocked conditions. Images are 200-µm longitudinal slices through the lymph node center. Scale bars represent 100 µm. The dashed line indicates the lymph node outline. Individual REX3 reporters and merge images show Cxcl9-RFP (magenta) and Cxc10-BFP (cyan). Images are representative of three individual experiments with at least four mice in each group in each experiment. (D and E) Analysis of Cxcl9-RFP and Cxcl10-BFP expression in DC subsets within peripheral lymph nodes of mice at d8 following challenge with either live or heat-inactivated (inactive) LCMV Armstrong. Half of each group received d0–1 treatments of IFNAR block. Data are representative of three individual experiments with three mice per group per experiment. Data are the mean ± SEM. (D) Graphs summarizing the graded frequencies of Cxcl9-RFP+ and Cxcl10-BFP+ DC subsets at d8 following acute LCMV Armstrong infection. (E) Frequency and total cell numbers and gMFI of chemokine reporter expression in each immune challenge condition. Each dot represents a single mouse. Data are the mean ± SEM. Statistical differences were analyzed using unpaired t tests. (F) Graphs summarizing the mean frequencies ± SEM of TEFF and TSCM cells in each condition. Data are representative of three independent experiments with three to four mice in each group. Statistical differences were analyzed using unpaired t tests. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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