Role of RAGE in acute arterial injury: studies in mice with genetically manipulated levels of RAGE/RAGE function. (a–f) RAGE null mice. Lung tissue retrieved from homozygous RAGE null mice or wild-type animals was subjected to RT-PCR for detection of RAGE mRNA or Western blotting (a and b). Mice were subjected to femoral artery guide-wire injury. I/M ratio was determined on day 28 (c) and van Gieson’s elastic staining was performed on a representative femoral artery section from a wild-type mouse (d) and a RAGE null mouse (e). Scale bar: 50 μm. (f) MPO activity. One hour after injury, vessel segments were retrieved from RAGE null and wild-type mice and subjected to MPO activity assays. Vessels were harvested and pooled from n = 2 mice per condition. Data from three sets of pooled animals per condition are reported. (g–l) Tg SM22-DN-RAGE mice. Tg SM22-DN-RAGE mice were identified by Southern blotting (g). RT-PCR was performed on samples from Tg mice overexpressing full-length RAGE (lanes 1 and 2) or DN-RAGE (lanes 3 and 4). (h) Western blotting was performed on lysates retrieved from the aortae of wild-type and Tg animals (i). Immunostaining with anti-RAGE IgG of femoral artery from Tg SM22-DN-RAGE mice (k) compared with a vessel from a non-Tg littermate (j). Scale bar: 25 μm. (l) Tg SM22-DN-RAGE mice and littermates were subjected to femoral artery injury and I/M ratio was determined on day 28. In l, there were at least 10–20 vessels per experimental condition.