Fig. 4.
B cell clusters form in the subarachnoid space adjacent to activated endothelial vasculature. Tam-BAPC mice were treated with Tam at week 3 post CD4 T cell transfer and killed 3 d post EAE onset. (A, Left) Decalcified spinal cords were stained for B220 (pink), CD3 (green), and DAPI (blue) and imaged at 4× magnification. (A, Right) Boxed insets are regions of interest (from left) stained for activated endothelial cells (VCAM-1, red), astrocytes (GFAP, green), and DAPI (blue) and imaged at 10× magnification. (B, Left, Top, and Bottom) Overlays of single-color images displayed at Right. (i) GFAP (green), (ii) VCAM-1 (red), (iii) DAPI (blue), and (iv) podoplanin-1 (white). (C) Serial sections of the spinal cord shown in B. (Left, Top, and Bottom) Overlay images of single-color images displayed at Right. (i) Laminin (green), (ii) B220 (red), (iii) DAPI (blue), and (iv) podoplanin-1 (white). All images are representative of n = 7 mice, pooled from at least 4 separate experiments. (Scale bars, 100 μm.)