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. 2003 Mar 28;100(8):4754–4759. doi: 10.1073/pnas.0730743100

Figure 1.

Figure 1

Intimal atherosclerotic plaque, media, and adventitial microvessels from female recipients (subjects 1–4, Table 1) who received a male BMT (XY → XX; A–H). (A) Hematoxylin and eosin staining showing a grade III atherosclerotic plaque from subject 1. L, lumen; filled arrowhead, internal elastic lamina. The two boxes indicate the subendothelial intima and the deep intima that are subsequently analyzed with dual α smooth muscle actin staining and FISH for Y chromosome in B and C. (B) Combined immunostaining for α smooth muscle actin and FISH for Y chromosome showing clusters of male cells (blue DAPI-stained nuclei with green dot) surrounded by positive α smooth muscle actin staining (red Cy-3 stain surrounding male nuclei, white arrows) in the subendothelial intima of plaque from subject 1. A row of white arrows point to a cluster of male actin-positive cells. Some actin staining cells were negative for the Y chromosome (open arrowhead). (C) Male smooth muscle cells deeper within the intima of plaque from subject 1 colabeling for α smooth muscle actin and the Y chromosome (white arrows). (D) Male smooth muscle cells in the media of coronary artery from subject 2 showing combined labeling for α smooth muscle actin and the Y chromosome (white arrows). The open arrowhead indicates the elastic lamellae of the media (M) and the filled arrowhead indicates the internal elastic lamina. (Inset) Chromosomal multiploidy analysis of male donor smooth muscle cells in female coronary artery showing diploid nature and lack of cell fusion. Shown are two separate nuclei (DAPI) each showing single X chromosome (large red dot) and Y chromosome (green dot) and also two chromosome 18 markers per nucleus (light blue dots, arrowheads), indicating each nucleus is diploid. The background red staining indicates α smooth muscle actin staining with Cy-3 fluorescence. (E and F) Male smooth muscle cells within the subendothelial and deep intima of a coronary artery plaque from subject 4 showing combined labeling for smooth muscle myosin heavy chain (E, red staining) and smooth muscle calponin (F, red staining) and the Y chromosome (white arrows). (G and H) Male smooth muscle cells in microvessels of the adventitia of an atherosclerotic coronary artery from subject 3 showing combined labeling for α smooth muscle actin and the Y chromosome (white arrows). The open arrowhead indicates an actin staining cell that was negative for the Y chromosome. (I) Combined FISH for X and Y chromsome and α smooth muscle actin costaining showing absence of male cells in a control diseased artery of a same-gender female BMT subject. In B–I the nuclei were counterstained with DAPI (blue).