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Figure 2 VSMC Proliferation in Collateral Arteries and Angiography of the Collateral Circulation
(A) Vascular smooth muscle cell (VSMC) proliferation in collateral arteries. In a double staining of alpha smooth muscle actin (green) and Ki67-protein for visualization of actively proliferating cells, the percentage of Ki67 positive nuclei (red) of all nuclei (blue) was quantified. Ki67 is a specific marker for proliferating cells, and only growing collateral arteries stain positive, whereas quiescent anastomoses from the non-occluded hindlimb remain negative. Collateral arteries from TGF-ß1–treated animals revealed a significantly higher index of vascular proliferation compared with the control groups, corresponding to the increase in functional conductance. (B) Angiography of the collateral circulation 7 days after stent implantation. After stent implantation in the external iliac artery (arrow), the site of vascular access in the superficial femoral artery was occluded by double ligation (*). Quantification of detectable collateral arteries with a stem, midzone, and re-entry demonstrated a weak trend toward a higher number of collaterals in the TGF-ß1–treated group. VSMC = vascular smooth muscle cell; other abbreviations as in Figure 1.
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