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Figure 4 | Acta Neuropathologica Communications

Figure 4

From: Intrastriatal injection of interleukin-1 beta triggers the formation of neuromyelitis optica-like lesions in NMO-IgG seropositive rats

Figure 4

The effects of cytokines and chemokines on rat brain endothelial cells in vitro . Rat brain endothelial cells (EC) were isolated from the cerebra of 3-week old (a,c) and adult (b,d) Lewis rats, and are shown here in phase-contrast microscopy (a,b) and after staining with an antibody against zonula occludens 1 (ZO-1, red; c, d; scale bar = 25 μm). (e) Endothelial cells from 3-week old (left panel) and adult (right panel) Lewis rats were stimulated for 22 hours with vehicle, IL-1β, IL-6, IFN-γ, TNF-α, or CXCL2, and then subjected to PCR analysis of the following genes: Cxcl1, Cxcl2, Csf3, Ccl2, Ccl5, Icam1, Vcam1, and GAPDH. These data are representative of 2 different, independently performed experiments. (f,g,h) The lysates of rat brain endothelial cells treated for 12 hours with IL-1β (f) or vehicle (g) were analyzed with antibody arrays and revealed changes in protein expression of CXCL1 (red rectangle), CXCL3 (gray rectangle) and CCL2 (green rectangle). The fold-change in signal intensity of these different proteins was then calculated using the Image J software of the National Institute of Health. (i,j) Rat brain endothelial cells cultured for 24 hours in the presence of IL-1β (i) reveal a marked increase in ICAM1 protein expression (green), which was not seen after 24 hours of culture in the presence of vehicle (j). All cells were counterstained with antibodies against ZO-1 (red). Bar = 25 μm. Data are representative of 2 different experiments.

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