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. 2017 Jul 17;7:5577. doi: 10.1038/s41598-017-06031-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Visualization of host lymphatic contribution in vascularized lymph node transfer model. (A) Superficial inferior epigastric artery (SIEA) free flaps were isolated from EGFP⁻ wild type Sprague-Dawley rats on a single vascular pedicle. (B) SIEA flap after transfer to a Prox1-EGFP recipient rat and completion of microsurgical anastomosis of the artery and vein. (C) Inset of the SIEA free flap using Prolene sutures, which facilitate identification upon harvest. (D) Gross appearance under a stereoscope of SIEA flap harvested after 30 days. Blue suture knot (arrow) marks the host-donor boundary. (E) EGFP signal at the inferior pole of the transferred flap at low magnification. (F) Enlarged image of the edge area demonstrating lymphatic vessels with EGFP⁺ host-derived LECs. (G–I) Pdpn immunofluorescence assay identified host contribution to lymphangiogenesis in the donor tissue. (J–L) Enlarged images of boxed areas in panels (G–I), respectively. Scale bars: 1 mm (D,E), 0.5 mm (F), 100 µm (G), 50 µm (J).