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Data Supplement for Arteriosclerosis, Thrombosis, and Vascular Biology: Volume 20, Issue 4 -- Page 957
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Data Files
- Figure I. Confirmation of high frequency of apoptosis by Taq polymerase-based in situ ligation assay. A, Positively-stained apoptotic cell is indicated with closed arrow that is surrounded by negatively-stained normal cells in human tonsil. B, In human umbilical vein at the same time of delivery, closed arrows indicate several apoptotic endothelial cells that are positively stained, whereas open arrow indictes negatively-stained endothelial cell that does not undergo apoptosis. C and D, In intima, as well as in media of human umbilical artery, apoptotic smooth muscle cells are frequently observed. Closed arrows indicate apoptotic cells; open arrow, normal cells. TS indicates tonsil; UV, umbilical vein; iUA, intima of umbilical artery; mUA, media of umbilical artery (bar=10 µm).
- Figure II. Transmission electron micrograph of vascular smooth muscle cells in the human umbilical artery at birth. A, Normal smooth muscle cells with endoplasmic reticulum and contractile filaments as dense bodies; B, smooth muscle cell undergoing apoptosis with cytoplasmic condensation and fragmentation of condensed nucleus; C, small apoptotic nuclei are indicated by closed arrows, whereas large normal smooth muscle cells is indicated by open arrow; D, closed arrows indicate several cells showing the early features of apoptosis, cytoplasmic condensation, and peripheral margination of condensed chromatin, in intima of umbilical atery where cells are arranged haphazardly. Acellular space indicated by triangle next to the apoptotic cells may be due to cell loss by apoptosis. Few normal-looking cells are indicated by open arrows. There is no inflammatory cell infiltration. (Double bar=3 µm in A, B, and C; single bar=7 µm in D.)
- Figure III. Transmission electron micrograph of endothelial cells in human umbilical vein at birth. A, Normal endothelial cell with elongated nucleus has broad base to contact with basal lamina. B, Endothelial cell undergoing apoptosis shrinks to a half-size of normal cell due to the condensed cytoplasm and nucleus. C, Closed arrows indicate 5 endothelial cells showing various stages of apoptosis, condensed cytoplasm, and nucleus; open arrows, two endothelial cells not showing apoptotic features yet. There is one apoptotic nucleus just beneath basal lamina. D, Closed arrow indicates apoptotic endothelial cell that shrinks due to condensation of cytoplasm and nucleus and loses its broad contact with basal lamina. Triangle indicates 3 endothelial cells that lose broad contact with basal lamina and shrink a little but do not show typical features of apoptosis yet. (Single bar=1.3 µm in A and B; double bar=3.5 µm in C and D.)
- Figure IV. Low-magnification view of hematoxylin and eosin staining of the ductus arteriosus with adjacent aorta and pulmonary artery (A) and aortic arch with its branching great arteries (B) from human neonates. Patterns of immunohistochemical and TUNEL staining were compared for ductus arteriosus and its adjacent arteries, and the aortic arch and the branching points of the great arteries on the same slide. Findings of rectangular areas specified in A or B were presented and compared as shown in print Figure 4 and online Figure V, respectively. Ao indicates aorta; CCA, common carotid artery; DA, ductus arteriosus; PA, pulmonary artery; and SCA, subclavian artery.
- Figure V.Comparison of the histological findings for the ductus arteriosus and the adjacent aorta in human neonates. A through D, Ductus arteriosus; E through H, aorta. A and E, Hematoxylin and eosin staining; B and F, TUNEL staining; immunohistochemical staining of Bax (C and G) and Bcl-2 (D and H). Pyknotic nuclei are observed in the ductus arteriosus as shown in panel A and are in contrast with the normal plump nuclei in the aorta, as shown in panel E. These pyknotic nuclei are TUNEL-positive as shown in panel B, whereas there is no positive cell in the aorta, as shown in panel F. This apoptosis of the ductus arteriosus is associated with the differential expression of Bax, which is expressed only in the ductus, not in the adjacent aorta, as shown in panels C and G. Bcl-2 is expressed neither in the ductus nor in the adjacent aorta, as shown in panels D and H. DA indicates ductus arteriosus; Ao, aorta. Bar=25 µm.
- Figure VI. Comparison of histological findings of the branching point of the the great artery and the aortic arch in human neonates. A through D, Branching point; E through F, aortic arch. A and E, Hematoxylin and eosing staining; B and F, TUNEL staining; immunohistochemical staining of Bax (C and G) and Bcl-2 (D and H). Pyknotic nuclei are observed in the branching point as shown in panel A and are in contrast with the normal plump nuclei in the aortic arch, as shown in panel E. These pyknotic nuclei are TUNEL-positive, as shown in panel B, whereas there is no positive cell in the aorta (panel F). This apoptosis of the branching point is associated with a differential expression of Bax, which is expressed neither in the branching point nor in the aortic arch, as shown in panels D and H. BR indicates branching point of great artery; Ao, aortic arch. Bar=25 µm.
Prepared by: the Data Supplement Manager
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