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. 2014 Apr 30;34(18):6316–6322. doi: 10.1523/JNEUROSCI.4912-13.2014

Figure 3.

Figure 3.

Peripheral macrophages are more susceptible than microglia to cell death after phagocytosis of myelin, in vitro. Fluorescent images showing BMDMs (Ai) and primary microglia (Bi) take up DiI-labeled myelin, in vitro. Aii, Bii, Representative FACS plots of CD45+ cells labeled for AnnexinV (apoptotic marker) and efluoro-780 (viability dye). Plots show level of apoptotic cell death (FITC-AnnexinV-positive, efluoro-780 viability dye positive; bottom right quadrant) and end-stage apoptosis/necrosis (efluoro-780 viability dye positive; top two quadrants) with increasing amounts of myelin added for 24 h. Aiii, Peripheral-derived macrophages show significant increase in apoptosis after treatment with 200 μg/ml myelin (24 h, 8.7-fold and 48 h, 6.1-fold). Biii, Primary microglia show much lower levels of apoptotic cell death in cultures treated with 200 μg/ml myelin (24 h, 1.7-fold and 48 h, 2.8-fold). End-stage apoptosis/necrosis increased 5.3-fold in peripherally derived macrophages at 24 h (200 μg/ml myelin), while necrotic cell death was not seen at either time point in primary microglia. Values expressed as mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.0001. n = 3–4. Scale bar, 20 μm.