Fig. 4

Colocalization of MIF and amyloid plaques in APP/PS mice. A, B Fixed brains were prepared for paraffin-embedded sectioning at 5 μm thickness. MIF was detected by anti-MIF antibody and visualized by ABC and DBA methods. Nuclei were stained by hematoxylin. The sections were observed under traditional microscopy. Arrows point to plaques associated MIF expression, which resembles the expression pattern of Aβ plaques. Scale bar, 100 μm. C Fixed brains were dehydrated in 30% sucrose solution and embedded in O.C.T. for cryosectioning at 30 μm thickness. MIF was detected by polyclonal MIF antibody, and Alexa 488-labeled secondary antibody, and Aβ plaques were detected by monoclonal 4G8 antibody and Alexa 594-labeled secondary antibody. Nuclei were stained with DAPI, and brain sections were observed under fluorescent microscopy. Arrowspoint to colocalization of MIF and plaques. Arrow heads point to MIF expression cells. Scale bar, 100 μm in B, and 25 μm in inserts and C