Figure 2.

Gal-9 modulates the recruitment of innate immune cells. Flow cytometry analysis was employed to determine in situ neutrophil and monocyte subsets. At the peak of inflammatory reaction (18h), ankle joints were digested, and single cell suspensions were obtained. Flow cytometry strategy applied to identify neutrophils (A), and monocytes (B), after Gal-9 treatment are shown. Cells were washed and stained with: CD45, LY6-C, LY6-G, CD11b, and CD115/CD45⁺ cells were plotted for LY6-C and LY6-G expression to identify CD45⁺/LY6-C^hi/LY6-G^hi as neutrophils (A). CD11b⁺ cells were plotted for LY6-C and CD115 expression to distinguish CD11b⁺/CD115⁺/LY6-C^lo patrolling monocytes from CD11b⁺/CD115⁺/LY6-C^hi inflammatory monocytes (B). (C) Total infiltrated leukocytes, (D) neutrophil and (E) inflammatory monocytes were quantified in the different experimental conditions. Representative FACS plots of three independent experiments with similar results are shown. Values are presented as means ± SEM of n = 6 mice per group. Statistical analysis was conducted by one-way ANOVA followed by Bonferroni’s for multiple comparisons. ^##P ≤ 0.01 vs Ctrl group; *P ≤ 0.05, **P ≤ 0.01 vs MSU group.