Brain Tissue Analysis in Nude Mice

Nude mice were euthanized, their brains were perfused and dissected as previously described [21 (link)], and 30-μm coronal sections were obtained. Free-floating sections of the brains from each group were washed three times with PBS and incubated at room temperature for 1 hour in PBS/Triton X-100 that contained 1% bovine serum albumin (BSA). Then, the sections were treated overnight with the primary antibodies of mouse anti human Ki67 and GFAP monoclonal antibody (Abcam, UK). After treatment with secondary biotinylated antibodies, color reactions were achieved with diaminobenzidine (DAB) (Sigma, USA) and counterstained with Mayer's hematoxylin.