In situ localization of OPN mRNA expression in the ischemic cortex. Shown is film autoradiography of OPN mRNA expression 5 d after permanent MCAO (A). OPN mRNA-expressing cells are located abundantly throughout the entire infarcted region. The ED-1 antibody and OPN mRNA colocalized in many, but not all, of the macrophages in the infarct (B). Morphologically, the ED-1-positive and OPN-positive cells (B,inset) have an ameboid appearance indicative of their activated phagocytic state. Scale bars: in B, 20 μm; in inset, 10 μm.