Fig. 4

Upregulation of MIF in ICV-STZ mice and colocalization with GFAP in the hippocampus. a Serial sections of WT mouse brain were stained for MIF protein using a rabbit anti-MIF polyclonal antibody and Alexa Fluor 488-conjugated anti-rabbit IgG (green fluorescence). The sections were subsequently stained for GFAP using a monoclonal anti-GFAP Cy3^TM antibody (red fluorescence). Nuclei were stained with DAPI (blue). Images with combined fluorescent channels are shown at ×200 (scale bar in the upper left panel 75 μm). Selected areas are enlarged by 10 times and shown as combined as well as individual fluorescence stains. Quantification of the MIF (b) and GFAP (c) fluorescence was shown after normalization of the green (MIF) and red (GFAP) fluorescence against the blue fluorescence (nuclei). Data shown are means ± SEM based on multiple experiments (n = 3). *p < 0.05, **p < 0.01 compared with WT mice receiving saline