Left: Rat cerebellum stained with mouse anti-MeCP2 (red, 1:500) and chicken anti-calbindin (C-1798-50, green, 1:2,000) by Immunohistochemistry. Blue: DAPI nuclear stain. IHC Method: Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 1 hour, cut to 45 um, and free-floating sections were stained. The MeCP2 antibody specifically labels nuclei of neuronal cells to a variable extent, while calbindin is expressed in the dendrites and perikarya of Purkinje cells in the molecular layer of cerebellum. Right: Western blot analysis with mouse anti-MeCP2 (green, 1:1,000) and chicken anti-GAP43 (red). (2) rat whole brain, (3) nuclear fraction of rat brain, (4) mouse whole brain, (5) nuclear fraction of mouse brain. A strong band at about 75 kDa is observed in rat and slightly lower in mouse nuclear enriched fractions, corresponding to MeCP2 protein. GAP43 protein is detected in all preparations with apparent molecular weight of 43 kDa.