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. 2010 Aug;58(8):721–730. doi: 10.1369/jhc.2010.955609

Figure 4.

Figure 4

Expression of nestin protein in neovascular cells in vivo. (A) In vivo neovascular branches grew for 7 days into an implanted silicone tube designed for a neoangiogenesis assay. Arrow indicates the open end of the tube where vascular ECs entered the tube. (B–D) Confocal images of neovascularization. (B) CD31 (red), (C) nestin (green), and (D) a merged image. Nestin was expressed in every CD31-positive EC in the tube. (E–H) Confocal images of neovascularization at the injury site of the mouse forebrain 14 days after cold injury. (E) CD31-positive (red) vasculature increased at the injury site. (F–H) The area denoted by a rectangle in E is shown at higher magnification to show staining for CD31 (F, red), nestin (G, green), and a merged image (H). An EC within the robust vasculature in the injured brain was double-positive for CD31 and nestin (arrows). A patchy perivascular lining of nestin-positive cells was also observed on the parenchymal border of CD31-positive ECs (arrowheads). (I–L) Images of the contralateral, uninjured side of the forebrain from the same mouse. (I) Fine, scattered CD31-positive (red) vasculature was observed in the uninjured mouse brain. (J–L) The area denoted by a rectangle in I is shown at higher magnification to show staining for CD31 (J, red), nestin (K, green), and coexpression in a merged image (L). Note that ECs in the normal brain did not express nestin. Bars: A = 1 mm; B–D,F–H,J–L = 20 μm; E,I = 50 μm.