Fig. 6

Hypothermia differentially modulates CD86 and CD206 expression in myeloid cells of the total hemisphere. Nine-day-old C57BL/6 mice were exposed to HI followed by 4 h HT or NT. Animals were sacrificed and analyzed via flow cytometry 1, 3 or 7 days after HI. CD11b⁺ cells were isolated via MACS from brains of uninjured sham-operated and HI-injured animals. Flow cytometry with a small aliquot of isolated cells was used to analyze the frequency of CD86⁺ (green gate), CD206⁺ (blue gate) and CD86/CD206⁺ cells (turquoise) cells (a, gates were set according to fluorescence minus one controls). Quantification of these subsets (b–d) was performed in macrophages (CD11b⁺CD45^high) and resident microglia (CD11b⁺CD45^int). *p < 0.05, **p < 0.01, ***p < 0.001, n = 6 animals/timepoint for sham (left and right hemisphere pooled), n = 12 animals/timepoint for HI + NT and for HI + HT (2 ipsilateral hemispheres pooled per sample). Contour plots in a are representative for cells isolated 7 days after HI