Figure 2: Sperm plasma induced the expression of IL-17A.

(a) IL-17A expression was analysed by quantitative PCR in uteri at the indicated days. The data are shown as the mean ± S.E.M. from four independent experiments and independent t-tests. (b) IL-17A expression was analysed by ELISA in uteri at the indicated times. The data are shown as the mean ± S.E.M. from four independent experiments and independent t-tests. (c) IL-17A expression was analysed by quantitative PCR in uteri from female mice that were mated with normal or vasectomized males on D0.5. The data are shown as the mean ± S.E.M. from five independent experiments with independent t-tests. (d) Uterine cells from virgin mice were stimulated with the Cell Stimulation Cocktail (eBioscience) and analysed for CD3, γδ TCR, IL-17A and CD45 expression in four independent experiments. The numbers represent the percentage of the population within the indicated gates. i: whole uteri cells; ii: gated on the IL-17A positive population; iii: gated on the CD3⁺ CD45⁺ population. (e) Uterine cells from virgin mice were co-cultured with different stimulators (Control: brefeldin A; Positive control: 2 μl/mL of the Cell Stimulation Cocktail; Sperm cell: sperm cell (1:1) and brefeldin A; Seminal plasma: 0.5% seminal plasma and brefeldin A) as indicated and analysed for CD3, γδTCR, IL-17A and CD45 expression after stimulation in four independent experiments. The representative FACS profile was gated by CD3⁺ CD45⁺. (f) T cells sorted from mouse uterine tissue was labelled with CD3, co-cultured with the different stimulators as indicated above and analysed for γδTCR and IL-17A expression in three independent experiments. The numbers represent the percentage of the population within the indicated gates.