Regulator of calcineurin 1 (Rcan1) expression colocalizes with glial fibrillary acidic protein (GFAP)-positive cells during transient focal brain ischemia. Coronal brain sections of animals subjected to 1 h middle cerebral artery occlusion (MCAO) followed by 24 h blood reperfusion. (A) Nissl staining revealing a hypochromatic area of ischemic tissue in the infarcted (I) neocortex. (B) Immunohistochemistry showing expression of GFAP protein (red) in the contralateral hemisphere (a) and the infarcted region (b). Nuclei are revealed by staining adjacent sections with TO-PRO 3 (blue; c and d). (C) Immunohistochemistry showing the GFAP-positive area around the infarcted brain region. The boxed region is magnified in the lower panels: (a) GFAP immunostaining (red); (b) TO-PRO 3 nuclear staining (blue); (c) Rcan1 immunostaining (green); (d) overlay of (a-c). Yellow indicates GFAP-Rcan1 colocalization.