Fig. 2

Analysis of microglia and myeloid immune cell subsets upon PACAP treatment. Immune cells were isolated from brain, blood and spleen, and analyzed by flow cytometry. Only single, live immune cells were considered for analysis. A Representative gating strategy applied for discrimination of CD45^hiCD11b⁻lymphocytes and CD45^intCD11b^int microglia (Mg). CD45^hiCD11b^hi myeloid cells (upper right gate) were further discriminated into Ly6G⁺ neutrophils, and Ly6G⁻ mononuclear cells were differentiated according to Ly6C expression in Ly6C^hi inflammatory monocytes, Ly6C^int monocyte-derived DCs and Ly6C^lo monocyte-derived macrophages. B, C Bar charts show total cell numbers of microglia and myeloid immune cell subsets in the brain. D, G Myeloid cells from blood and spleen were first identified as CD11b⁺ (not shown). Contour plots show the classification of selected cells into monocyte subsets (according to Ly6C expression) and Ly6G⁺ neutrophils. Bar charts compare the cell frequency in the blood (E, F) and spleen samples (H, I) as percentages from parent population. Data show individual values as mean + SEM, n = 4–5, *p < 0.05, **p < 0.01 (unpaired two-tailed t-test) from a representative experiment; Control (black bars) and PACAP-treated (white bars)