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. 2017 Jun 19;135(25):2505–2523. doi: 10.1161/CIRCULATIONAHA.116.025722

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© 2017 The Authors.

Circulation is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 5. — SOX17 is required for the induction of induced endothelial cells (iECs) from CD34⁺ progenitors. A, Heat map of endothelial developmental genes during iEC induction (iECs at days 0, 3, 7, and 10) by quantitative real-time polymerase chain reaction (PCR). The color key indicates gene expression level relative to B2M (beta-2-microglobulin) normalized to iECs at day 0. Blue to red color represents low to high expression level. B and C, The expression level of SOX17 in iECs. The time course of mRNA level of SOX17 during the induction of iECs (n=4). B, Immunoblot of SOX17 in adult fibroblasts and iECs (C). D through F, The representative immunoblots (D) and their statistical analysis (E), and the quantitative PCR analysis (F) of SOX17 and EC-specific markers VE-cadherin, CD31, and FLK1 in iECs infected with lentivirus-expressing SOX17-shRNA or scramble shRNA (n=4). G–I, The representative immunoblots (G) and their statistical analysis (H), and the quantitative PCR analysis (I) of SOX17 and EC-specific markers VE-cadherin, CD31, and FLK1 in iECs infected with retrovirus-expressing SOX17-GFP or GFP (n=4). J and K, Representative flow-cytometry overlay plots of the endothelial markers VE-cadherin (J) and statistical analysis (K) of percentage of VE-cadherin–positive cells in iECs overexpressed with SOX17-GFP or GFP. Data are presented as mean±SE; *P<0.05, ***P<0.001, compared with day 0 in A; ^#P<0.05, ^###P<0.001, compared with day 3 in A; ^&P<0.05 compared with day 7 in A. *P<0.05, **P<0.01, compared with control in E, F, H, and I. Using repeated measures 1-way analysis of variance in B, the Student t test with multiple comparisons (Bonferroni Correction) in E, F, H, and I, and the Student t test in K.