Relevance of GlcNAc and CD206 during engulfment. (A) Control Th17-skewed cells and GlcNAc-pretreated Th17-skewed cells were co-cultured with organotypic hippocampal slices. Co-cultures of organotypic hippocampal slices from CX3CR1^GFP pups with pathogenic B6.2D2.CFP.Th17 cells were stained with Image-iT LIVE Red Caspase-3 and -7 Detection Kit to identify apoptotic cells and imaged over a time period of 20 min. Only apoptotic T cells were considered for further analyses. Mean (± SEM) percentages of engulfment rate among the interactions of GlcNAc-pretreated Th17-skewed cells in organotypic hippocampal slices (n = 4 organotypic slices from three different experiments) normalized to the GlcNAc-untreated control group (n = 4 organotypic slices from three different experiments). Statistical analysis was performed using two-sided Student’s t tests. (B) Representative visualization of CD31 (magenta), CD206 (red), CX3CR1 (green), and DAPI (blue), in dexamethasone/LPS/IL-4–treated organotypic hippocampal slice cultures (representative for n = 4 reslices from two different experiments). Scale bar = 20 µm. Arrows point to CD206⁺ myeloid cells. (C) Representative staining of DAPI (blue), CD206 (red), CD4 (magenta), and Iba-1 (green) in EAE diseased animals (n = 3 mice [C57BL/6 or Cx3CR1.GFP] from two different experiments). The second panel demonstrates a probable blood vessel. Magnification of a slice view of CD206⁺ engulfing parenchymal (pc) and perivascular (pv) myeloid cell is shown in the third and fourth panel. Scale bars = 30 µm in first and second panel and 20 µm in the third and fourth panels.