Fig. 3

Double immunohistochemistry confirms Rab32 staining in microglia/macrophages and axons in MS. Rab32 positive staining (brown) was investigated in microglia/macrophages (CD68, blue/gray) and axons (neurofilament, blue/gray) with co-localization producing a black stain. a Staining of MS NAWM; Rab32 (brown) staining of CD68-positive (+ve) microglia cells. b Rab32 expression was also observed in microvascular cells (arrow). c Oil red-O staining of an acute MS lesion. The dashed line and small arrows depict the lesion border, and red staining shows myelin ingestion by macrophages. The large arrows show the blood vessels. The asterisk depicts surrounding NAWM. d Rab32-positive (+ve) microglia in the acute lesion border and Rab32-negative (−ve) microglial cells in adjacent NAWM. The dashed lines represent the area shown at higher power. (see high-magnification insets E, F, and G). e Rab32-negative (−ve) CD68+ve microglial cells in NAWM. h, i Staining of acute lesion tissue labeled for Rab32 (brown), macrophages (blue/gray, h), and axons (blue/gray, i). f High magnification of CD68-positive (+ve) microglial cells staining for Rab32 in acute lesions. g High power magnification of axons (blue/gray, co-localized with Rab32 as black), in close proximity to Rab32-positive (+ve) macrophages (brown). Scale bars in a and b = 25 μm; c, d, h, and i = 50 μm; and 12.5 μm in e, f, and g