Figure 3

UP-derived CD4⁺CXCR5^hiPD-1^hi Tfh cells show high levels of Tfh-associated molecules and display a memory/activation phenotype.

(a) CD4⁺ T cells were gated from the lymphocytes in the UP of pregnant mice on E11.5, and a schematic diagram illustrates the strategy used to analyse the molecular expressions of the indicated proteins among CD4⁺CXCR5^loPD-1^lo, CD4⁺CXCR5^hiPD-1^lo, CD4⁺CXCR5^loPD-1^hi and CD4⁺CXCR5^hiPD-1^hi cells. (b) Comparisons of CXCR5, PD-1, ICOS and CD44 expression among the indicated CD4⁺ T-cell subsets derived from the UP on E11.5. (c,d) Intra-nuclear expression of BCL-6 was detected according to the Foxp3 staining protocol, and representative flow histograms and cumulative data illustrate BCL-6 level within CD4⁺CXCR5^loPD-1^lo and CD4⁺CXCR5^hiPD-1^hi cells derived from the UP of pregnant mice on E11.5. Each symbol reflects the data from a single mouse (n ≥ 4 mice per group) and the data are representative of at least three independent experiments. Geometric MFI values were calculated using FlowJo 7.6.1 software and the data were assessed statistically using one-way ANOVA followed by Tukey’s post-tests (b) or Wilcoxon matched pairs test (d). UP: uterus and placenta; MFI: mean fluorescent intensity; hi: high; lo, low; **p < 0.05; ***p < 0.001.