Immigrant macrophages mediate the detrimental effect of RAGE on the infarct size. A, Bone marrow of ACTB–EGFP mice was transplanted to wild-type mice. Six weeks after transplantation, MCAO was performed. Two days later, FACS analysis showed that GFP+/CD45+ cells increased in the ischemic hemisphere compared with the nonischemic contralateral hemisphere (top row). GFP+ cells (bottom row) were mainly CD45hi/CD11+ in the ischemic hemisphere. B, Immunohistochemistry of Iba1 showed colocalization of EGFP and Iba1. Scale bar, 50 μm. C, Detection of RAGE and cyclophilin mRNA in blood by real-time RT-PCR confirmed the efficiency of bone-marrow transplantation. After PCR, products were resolved on an agarose gel next to a DNA marker (top, 100 bp marker; bottom, 25 bp marker). For quantification, see Materials and Methods. WT>WT, Transplantation of wild-type bone marrow to wild-type mice. RAGE−/−>WT, Transplantation of RAGE−/− bone marrow to wild-type mice. D, Immunohistochemical double-staining of RAGE(Cy3,red) and the microglial marker Iba1 [FITC (fluorescein isothiocyanate), green] in WT>WT and RAGE−/−>WT brains. Arrows, Double-labeled cells. Scale bar, 50 μm. E, Infarct volume in WT>WT was larger than in RAGE−/−>WT mice 48 h after MCAO. *p < 0.05 (n = 8–10, t test). F, Distribution of infarcts on coronal sections.