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. 2022 Jul 28;131(5):424–441. doi: 10.1161/CIRCRESAHA.122.321187

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© 2022 The Authors.

Circulation Research 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 2. — UCP2 (Uncoupling protein 2) is a target of KLF2 (Krüppel-like factor 2). A, Real-time quantitative polymerase chain reaction (qPCR) data showing KLF2 overexpression by adenovirus KLF2 (Ad-KLF2) reversed oscillatory shear stress (OSS)-induced downregulation of UCP2 mRNA in human umbilical vein ECs (HUVECs), OSS time is 12 h, n=5, 2-way ANOVA, Tukey post hoc. B, Real-time qPCR data showing KLF2 knockdown by KLF2-shRNA abolished unidirectional shear stress (USS)-induced upregulation of UCP2 mRNA in HUVECs, USS time is 24 h, n=5, 2-way ANOVA, Tukey post hoc. C, Western blotting data showing Ad-KLF2-mediated KLF2 overexpression for 24 h induces UCP2 protein expression in HUVECs. D, Ad-KLF2 overexpression for 24 h induces UCP2 mRNA expression in HUVECs and human aortic ECs (HAECs). n=5, nonparametric Mann-Whitney test. E, Endothelium-specific overexpression of Klf2 mediated by Ad-Cdh5-Klf2 in C57 mice for 7 d upregulated mRNA levels of Klf2, Ucp2, and Nos3 in mouse aortas. n=5, nonparametric Mann-Whitney test. Representative western blots and summarized data (F) showing that endothelium-specific overexpression of Klf2 mediated by Ad-Cdh5-Klf2 in C57 mice for 7 d upregulated UCP2 protein expression in mouse aortas. n=5, nonparametric Mann-Whitney test. KLF2 knockdown by KLF2-shRNA downregulated mRNA (G) and protein (H) levels of KLF2, UCP2, and eNOS (endothelial nitric oxide synthase) in HUVECs. n=6, nonparametric Mann-Whitney test. I, Dual luciferase activity assay results showing KLF2 regulates UCP2 promoter activity in HEK293A cells transfected with different lengths of human UCP2 promoters. n=4, 2-way ANOVA, Tukey post hoc. J, KLF2 increases UCP2 promoter activity in HUVECs electroporated a −508 bp UCP2 promoter. n=4, nonparametric Mann-Whitney test. K, Chromatin immunoprecipitation (ChIP)-qPCR assay in HUVECs transduced with Ad-KLF2 showing enriched KLF2 binding to UCP2 promoter. n=4, nonparametric Mann-Whitney test. Representative images were selected as they best represent images obtained. SCR indicates scramble.