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. 2019 Nov 6;9:16130. doi: 10.1038/s41598-019-52513-x

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© The Author(s) 2019

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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Isolation and characterization of EPCs. BM-EPCs were isolated by density gradient centrifugation and were cultured until they reached the appropriate density. Isolated EPCs were grouped and incubated with FITC-labelled primary antibodies against the surface markers of EPCs, such as CD34, CD133, FLK-1, and vWF. Flow cytometry analysis showed that there were FITC-positive cells with specific EPC surface markers, such as (A) CD34, (B) CD133, (C) FLK-1 and (D) vWF. Isolated EPCs that were not incubated with FITC-labelled antibodies were tested as a control. The experimental group is marked in red, and the control group is marked in blue. Both the FITC-positive and FITC-negative cell percentages were calculated and are shown in the image. (E) Representative images of the FITC-UEA-I and Dil-Ac-LDL dual-staining of EPCs. Cell nuclei were stained with DAPI (blue fluorescence), FITC-UEA-I is shown in green and Dil-Ac-LDL is shown in red. The merged picture shows the overlay of the results for both FITC-UEA-I and Dil-Ac-LDL staining, showing dual-staining positive cells, which were characterized as EPCs.