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Immunoperoxidase staining of mouse embryos was done with monoclonal antibodies to PECAM (Pharmingen, San Diego, CA), endoglin (Pharmingen), FLK-1 (Santa Cruz Biotechnology, Santa Cruz, CA), or α-smc actin (clone 1A4, 1:500; Sigma, St. Louis, MO). Staining was developed in 3,3′-diaminobenzidine chromagen (Vector Laboratories, Burlingame, CA). Sections of stained tissue were counterstained with eosin B.
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Tissue was fixed in 3% glutaraldehyde and sequentially stained with osmium tetroxide, tannic acid, and uranyl acetate. After dehydration, tissue was embedded in Epon. Thin sections (60 nm) were counter-stained with uranyl acetate and lead citrate and examined on a JEOL 1200 electron microscope.
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Supported by National Institutes of Health grants K08 HL03490-03 and T35 HL07744-06, the Culpeper Scholarship in Medical Science, and a University of Utah Seed Grant. We thank M. T. Keating for guidance and support and J. Miano and K. Thomas for clones and discussion. B.S.B. is a Howard Hughes Medical Institute Medical Student Research Training Fellow
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Supported by National Institutes of Health grants K08 HL03490-03 and T35 HL07744-06, the Culpeper Scholarship in Medical Science, and a University of Utah Seed Grant. We thank M. T. Keating for guidance and support and J. Miano and K. Thomas for clones and discussion. B.S.B. is a Howard Hughes Medical Institute Medical Student Research Training Fellow.
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