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Fig. 1 | Molecular Neurodegeneration

Fig. 1

From: Quantitative proteomics of acutely-isolated mouse microglia identifies novel immune Alzheimer’s disease-related proteins

Fig. 1

Study design and analytic approach for comprehensive quantitative proteomic analysis of murine microglia. a Work-flow summarizing isolation and purification of acutely isolated CD11b+ CNS immune cells from untreated 6–7 mo WT (C57BL/6 J) mice, LPS-treated WT mice (4 daily i.p. doses, 20 μg/dose) and age/sex-matched 5xFAD transgenic mice were (N = 3 pools per group, 2 brains per pool). Following percoll density centrifugation, cells were enriched for CD11b+ cells by MACS magnetic separation. b Flow cytometric confirmation of successful enrichment of CD11b+ cells after MACS enrichment. Pre-enrichment (top), CD11b+ cells account for 50–60% of all live CNS mononuclear cells. Post-enrichment (bottom), CD11b+ cells account for > 95% of all live cells. c Flow cytometric confirmation of minimal presence of peripherally derived CD11b + CD45high CNS-infiltrating macrophages or perivascular macrophages in untreated WT, LPS-treated WT and 5xFAD mice. d Proteomic work-flow for tandem mass tag (TMT) mass spectrometry based quantification. e Analytic approach used for differential expression analyses of datasets

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