Detection and quantification of neutrophils and monocytes in the total lower FRT during the estrous cycle by flow cytometry.

<p>Lower FRT was isolated from naïve adult virgin female C57BL/6 mice and prepared for flow cytometry. Cells were gated as CD45.2⁺CD11b⁺CD11c^lo (as shown in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114824#pone.0114824.s001" target="_blank">Figure S1</a></b>) prior to further analysis. (<b>A</b>) Percentage of CD45.2⁺CD11b⁺CD11c^loLy6C⁺Ly6G⁺ cells (neutrophils) and CD45.2⁺CD11b⁺CD11c^loLy6C⁺Ly6G⁻ (monocytes) at each cycle stage (estrus,E; proestrus, PE: metestrus, M; diestrus, DE). (<b>B–D</b>) Frequency (<b>B</b>), absolute number (<b>C</b>) and morphology (<b>D</b>) of neutrophils at each cycle stage. (<b>E–G</b>) Frequency (<b>E</b>), absolute number (<b>F</b>) and morphology (<b>G</b>) of monocytes at each cycle stage. Samples in <b>(D)</b> and <b>(G)</b> were Giemsa stained cytospins after sorting. Representative images shown in <b>(D)</b> and <b>(G)</b> were taken at 400× magnification. Data derived from three independent flow cytometry experiments (each run with tissue pooled from four mice per time point) were quantified and analyzed using one-way ANOVA with Tukey's post-test. The mean±SEM is shown (*, p≤0.05; **, p≤0.01; ***, p≤0.001).