Figure S3.
Analysis of Prdm1- and Zc3h12c-deficient mice during N. brasiliensis infection. Related to Fig. 4. (A) Gating strategy for lymphocytes shown in Fig. 4 A and Fig. S3, C and E. (B) Gating strategy of myeloid cells shown in Fig. 4 C and in Fig. S3, C and E. (C) Quantification of flow cytometric analysis of lung and mLN ILC2s and eosinophils (Eos) from untreated and infected Zc3h12cfl/fl and infected Nmur1CreZc3h12cfl/fl mice. Worm burden of untreated and infected mice. Mice were infected with N. brasiliensis for 7 days. Data are representative of two independent experiments; n = 4–5 mice per group. (D) Flow cytometric plots of ST2 expression (related to Fig. 4 B) in mLN and lung ILC2s from infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice; n = 4–6 mice per group. (E) Quantification of flow cytometric analysis of lymphocytes and myeloid cells from naive and infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice. Data are representative of two independent experiments; n = 3–6 mice per group. (F) Gating strategy for G. (G) Flow cytometric analysis of the expression of transcription factors GATA3 and cMaf in ILC2s from untreated and infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice (n = 3–5 mice per group). (H) Relative expression of Il9r in lung ILC2s from untreated and infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice. Mice were infected with N. brasiliensis for 7 days (n = 4–5 mice per group). N. b., N. brasiliensis. Mean ± SD; Student’s t test (H) or one-way ANOVA (C, E, and G); ns, nonsignificant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Refer to the image caption for details. Panel A shows the gating strategy for lung lymphocytes. Sequential gating identifies CD45 plus cells and distinguishes B cells, CD8 T cells, CD4 T cells, NK cells, ILC2s (Lin- CD127+ ST2+ KLRG1+), and ILC3s. Panel B shows the gating strategy for lung myeloid cells, identifying eosinophils, neutrophils, dendritic cells, and macrophages based on lineage and surface markers. Panel C presents flow cytometric analysis of lung and mLN ILC2 s and eosinophils in naive and N. b. infected Zc3h12c fl/fl and Nmur1 Cre Zc3h12c fl/fl mice, along with worm counts, showing altered immune responses in conditional knockout mice. Panel D shows representative flow cytometry plots of ST2 expression on mLN and lung ILC2s from infected Prdm1 fl/fl and Nmur1 Cre Prdm1 fl/fl mice. Panel E quantifies flow cytometric analysis of ILC2 s, total lymphocytes, and myeloid cells in lung and mLN from naive and infected Prm1 fl/fl and Nmur1 Cre Prdm1 fl/fl mice. Panel F shows the gating strategy for intracellular transcription factor staining, identifying GATA3 and cMaf expression in ILC2s. Panel G presents flow cytometric analysis of GATA3 and cMaf expression in lung and mLN ILC2s from naive and infected P r d m 1 f l slash f l and Nmur1 Cre Prdm1 fl/fl mice. Panel H shows relative Il9r expression in lung ILC2s under the indicated conditions.

Analysis of Prdm1 - and Zc3h12c -deficient mice during N. brasiliensis infection . Related to Fig. 4. (A) Gating strategy for lymphocytes shown in Fig. 4 A and Fig. S3, C and E. (B) Gating strategy of myeloid cells shown in Fig. 4 C and in Fig. S3, C and E. (C) Quantification of flow cytometric analysis of lung and mLN ILC2s and eosinophils (Eos) from untreated and infected Zc3h12cfl/fl and infected Nmur1CreZc3h12cfl/fl mice. Worm burden of untreated and infected mice. Mice were infected with N. brasiliensis for 7 days. Data are representative of two independent experiments; n = 4–5 mice per group. (D) Flow cytometric plots of ST2 expression (related to Fig. 4 B) in mLN and lung ILC2s from infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice; n = 4–6 mice per group. (E) Quantification of flow cytometric analysis of lymphocytes and myeloid cells from naive and infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice. Data are representative of two independent experiments; n = 3–6 mice per group. (F) Gating strategy for G. (G) Flow cytometric analysis of the expression of transcription factors GATA3 and cMaf in ILC2s from untreated and infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice (n = 3–5 mice per group). (H) Relative expression of Il9r in lung ILC2s from untreated and infected Prdm1fl/fl and infected Nmur1CrePrdm1fl/fl mice. Mice were infected with N. brasiliensis for 7 days (n = 4–5 mice per group). N. b., N. brasiliensis. Mean ± SD; Student’s t test (H) or one-way ANOVA (C, E, and G); ns, nonsignificant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

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