Mechanisms underlying sRAGE-mediated suppression of neointimal expansion in wild-type mice. (a) BrdU-labeling index. Mice were subjected to femoral artery guide-wire injury followed by intraperitoneal injection of BrdU and sacrifice on day 7. Nuclei immunoreactive with BrdU were quantitated, and the index in the neointima was reported. (b) TUNEL index. Mice were subjected to femoral artery guide-wire injury followed by sacrifice on day 7. Vessels were subjected to TUNEL staining, and data are expressed as an index of TUNEL-positive nuclei in the neointima. (c and d) Activation of Erk1/2 and PKB. Mice were subjected to femoral artery guide-wire injury and sacrificed 30 min later. SDS-PAGE/Western blotting with Ab’s to phospho-Erk1/2 (p-Erk1/2) or total Erk1/2 (c) or phospho-PKB (p-PKB) or total PKB were employed. (e and f) Activation of Jak2/Stat3. Mice were subjected to femoral artery guide-wire injury and sacrificed 7 days later. SDS-PAGE/Western blotting with Ab’s to phospho-Jak2 (p-Jak2) or total Jak2 (e) or phospho-Stat3 (p-Stat3) or total Stat3 (f) were employed. (g) Transcripts for MMP12, tenascin-C, and β-actin. Mice were subjected to femoral artery guide-wire injury. On day 7, total RNA was harvested for RT-PCR using the indicated primers. (h, i, and j) Masson’s trichrome stain. Mice were subjected to femoral artery guide-wire injury and sacrificed on day 28. Sections of femoral artery were stained with Masson’s trichrome reagent (i and j), and images were analyzed to determine the area occupied by blue staining ECM (h). Scale bar: 50 μm. The above experiments employed at least eight vessels for each experimental condition, and experiments were repeated three times.