Figure 1. Nestin-GFP⁺ cell population at the arterial remodelling sites represents a subset of MSCs.

Nestin-GFP mice were subjected to either sham surgery (Sham) or wire insertion-induced injury in femoral arteries (Injury). Blood samples were collected 1 week(1W) later. Percentages of blood CD45⁻ GFP⁺ cells out of total mononucleate cells from the mice were analysed by flow cytometry (a). n=5 mice per group. Data are represented as mean±s.e.m. *P<0.001 as determined by Student's t-tests. CD45⁻GFP⁺ cells were FACS sorted from circulating blood of the arterial injured mice (b, left panel). Representative image showing the percentage of the CD45⁻GFP⁺ cells that express LepR (b, right panel). The percentages of the GFP⁺LepR⁺ cells (c) or GFP⁺LepR⁻ cells (d) that express Sca1, CD105 or CD31, respectively, were analysed by flow cytometry. The data represent mean±s.e.m. from three to five mice from at least three independent experiments. Nestin-GFP mice were subjected to either sham surgery (Sham) or wire insertion-induced injury in femoral arteries (Injury). At 1W after the surgery, femoral arteries were harvested. Haematoxylin and eosin (H&E) staining of mouse femoral artery sections from the mice (e). A, adventitia layer; I, intima layer; M, media smooth muscle layer. Double-immunofluorescence images of femoral artery tissue sections from the mice using antibodies against GFP (green) and either LepR (f), αSMA (g), CD11b (h) or CD31 (i), respectively. *E, elastic fibre. White arrows represent double positive staining cells. 4,6-Diamidino-2-phenylindole stains nuclei blue. Scale bars, 50 μm. Quantification of the percentage of GFP⁺ cells that express LepR, αSMA, CD11b or CD31 at the injured sites (j). n=5. Data are represented as mean±s.e.m.