Journal: PLoS ONE

Article Title: Ancestral Vascular Lumen Formation via Basal Cell Surfaces

doi: 10.1371/journal.pone.0004132

Figure Lengend Snippet: (A) Confocal image of a transverse section through an anterior part of juvenile amphioxus stained for laminin (red) to localize basement membranes, and acetylated tubulin (green) to localize apical cilia. Cell nuclei are stained with DAPI (blue). NT, neural tube. NC, notochord. MYO, myomere. CO, coelom. Scale bar, 20 µm. (B) Higher magnification of the boxed area outlined in (A) shows laminin (red) lining the lumen (asterisk) of the amphioxus aorta. The apical cilia (green) of the intestinal cells project into the intestinal lumen. Nuclei are labeled with DAPI (blue). Scale bar, 5 µm. (C) The red channel from (B) shows laminin staining (arrow) in the lumen (asterisk) of the amphioxus aorta. Scale bar, 5 µm. (D) Confocal image of a transverse section through an E9.0 mouse embryo shows a dorsal aorta with a lumen (asterisk). Endothelial cells stained for CD31 (green) line the vascular lumen and separate it from the laminin-containing basement membrane (red) of the intestinal epithelium. Nuclei are labeled with DAPI (blue). Scale bar, 20 µm. (E) Higher magnification of the boxed area outlined in (D) shows endothelial cells (green) lining the aortic lumen (asterisk) with their laminin-free luminal cell surface (arrow). The abluminal cell surface (arrowhead) is adjacent to the laminin-containing basement membrane of the intestinal epithelium (red). DAPI (blue). Scale bar, 10 µm. (F) The red channel from (E) shows laminin staining on the abluminal cell surface (arrowhead) of the mouse aorta. Scale bar, 10 µm.

Article Snippet: The following primary antibodies were used: rat anti-mouse CD31 (PECAM-1) (BD Pharmingen), rat anti-mouse β1-integrin and rabbit anti-mouse collagen IV (Chemicon), mouse anti-β1 integrin (Santa Cruz), rabbit anti-laminin-111 (EHS) (Sigma), rabbit anti-laminin chain α1 , α4, and α5 , goat anti-podocalyxin (R&D), mouse anti-acetylated tubulin (Sigma).

Techniques: Staining, Labeling

Journal: PLoS ONE

Article Title: Ancestral Vascular Lumen Formation via Basal Cell Surfaces

doi: 10.1371/journal.pone.0004132

Figure Lengend Snippet: (A) Confocal image of a tubular network formed by Mile Sven 1 (MS1) cells upon Matrigel overlay. Vessels are stained for CD31 (green). Scale bar, 200 µm. (B) Confocal image at higher magnification of a vessel stained for CD31 (green) and cell nuclei (DAPI, blue). The lumen is labeled with asterisks. Scale bar, 10 µm. (C) Phase-contrast image of a hematoxylin-eosin stained vessel with a lumen (asterisks). Scale bar, 10 µm. (D) Electron micrograph showing a cross-section through a vessel with a lumen (asterisk). Microvilli (ˆ), characteristic of apical cell surfaces, are exclusively localized on the abluminal plasma membrane, whereas the luminal plasma membrane has a smooth appearance, characteristic of basal cell surfaces. Scale bar, 2 µm.

Article Snippet: The following primary antibodies were used: rat anti-mouse CD31 (PECAM-1) (BD Pharmingen), rat anti-mouse β1-integrin and rabbit anti-mouse collagen IV (Chemicon), mouse anti-β1 integrin (Santa Cruz), rabbit anti-laminin-111 (EHS) (Sigma), rabbit anti-laminin chain α1 , α4, and α5 , goat anti-podocalyxin (R&D), mouse anti-acetylated tubulin (Sigma).

Techniques: Staining, Labeling