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. 2020 Aug 12;11:1946. doi: 10.3389/fimmu.2020.01946

FIGURE 1.

FIGURE 1

Effects of CLP on total B cells and B cell subtypes in the spleen. C57Bl/6 laboratory mice underwent CLP and were euthanized at given timepoints. Data obtained using flow cytometry. Unmanipulated mice were used as T0 controls. Each point represents results in an individual animal, central horizontal line indicates mean, upright, and inverted Ts indicate standard deviation, data representative of two independent experiments. *p < 0.05, **p < 0.01 for ** relative to T0 using one-way ANOVA with Dunnett post hoc correction for multiple comparisons. (A) Total splenic B cells at given time post-CLP. Gating: FSC/SSC, singlets, Live, CD19+/B220+; N = 4–6/group. (B) Total mature (left) and immature (right) splenic B cells at given time post-CLP. Gating: FSC/SSC, singlets, Live, CD19+/B220+, CD93; N = 4–6/group. (C) Total follicular, marginal zone, and germinal center B cells per spleen at given time post-CLP. Gating: Follicular B cells: FSC/SSC, singlets, Live, CD19+/B220+, CD93–, B220+/CD138–, IgMlo/CD21/35lo; Germinal center: FSC/SSC, singlets, Live, CD19+/B220+, CD93–, B220+/CD138–, IgMlo/CD21/35lo, GL7+; Marginal zone: FSC/SSC, singlets, Live, CD19+/B220+, CD93–, B220+/CD138–, IgMhi/CD21/35hi; N = 5/group for Follicular, 3–4/group for germinal center B cells, 4–10 for marginal zone. (D) Total T1 transitional immature B cells per spleen at given time post-CLP. Gating: FSC/SSC, singlets, Live, CD19+/B220+, CD93+, IgMhi/CD23–; N = 3–4/group. (E) Total plasma cells per spleen at given time post-CLP. Gating: FSC/SSC, singlets, Live, CD19+/B220+, CD93–, B220+/CD138+; N = 3–4/group. (F) Total splenic CD4 T follicular helpers in the spleen at given time post-CLP. Gating: FSC/SSC, singlets, Live, CD90+/CD4+, PD1+/CXCR5+; N = 3–4/group.