Figure 2: Flow cytometric analysis of meninges of mice three days after SAH with the RORγt reverse agonist GSK805 (ROR inh) or vehicle.

A) Shows representative images of single cell suspensions of meninges. Samples were gated on singlet, live/dead, and CD45⁺ and the image shows a flow plot with CD11b staining on Y axis and Ly6G staining on X axis. The images show that there are two predominant myeloid cell types after SAH: monocytes (CD45⁺Ly6C^highLy6G^intCD11b^high/low), and neutrophils (CD45⁺Ly6C^lowLy6G^highCD11b^high). Monocytes can be classified as either CD11b^high or CD11b^low. B) There is a significant difference between the percentage of neutrophils in the vehicle treatment SAH animal and the GSK 805 (ROR inh) SAH animals (p=0.03) suggesting an effect of GSK805 on neutrophil entry. There were no differences between the CD11b^low or CD11^high monocyte groups (see Supplemental Figure 1B). The absolute number of neutrophils in the meninges of both groups was not different, likely due to retained inflammatory cells adherent to the walls of blood vessels. To investigate this, we proceeded to immunohistochemical evaluation (Figure 3).