RAGE blockade and neointimal expansion after acute arterial injury. (a–c) Femoral artery guide-wire injury was performed, and animals were followed until the indicated day, during which time they were treated with either sRAGE (100 μg/day, intraperatoneally; black bars) or MSA (100 μg/day; white bars) from days 0 to 7 (day 1 is the day of injury). I/M was determined (a) or vessels were stained with van Gieson’s elastic stain. (b) MSA. (c) sRAGE. Scale bar: 50 μm. (d) Animals were treated with the indicated dose of sRAGE or MSA followed by harvest of the vessels on day 28, and I/M ratio was determined. (e) Animals were subjected to femoral artery injury and then treated with sRAGE over the indicated interval. At 28 days, animals were sacrificed and I/M ratio determined. (f) Animals were subjected to femoral artery injury and received F(ab′)₂ (100 μg/day, intraperitoneally; days 0–7) prepared from either nonimmune rabbit IgG or rabbit anti-RAGE IgG. (g) Serum from sRAGE-treated mice subjected to femoral artery injury was immunoprecipitated with anti-AGE IgG; immune precipitate was solubilized and subjected to Western blotting with anti-RAGE IgG. Lane 1, MSA treatment from days 0 to 3; lane 2, sRAGE from days 0 to 3; lane 3, normal mouse serum without injury; lane 4, sRAGE treatment and immunoprecipitation with nonimmune IgG. In (a–f) at least 10 to 20 vessels per condition were analyzed by an investigator blinded to the experimental conditions. The immunoprecipitation study (g) was performed three times.