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. 2020 Jan 21;10:842. doi: 10.1038/s41598-020-57673-9

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© The Author(s) 2020

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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CRIF1 deficiency mediated BH4 biosynthesis diminution in HUVECs. (A) Schematic representation of the BH4 de novo pathway and recycling pathway. (B) Fluorescent signals of total biopterin and BH₂ + B by fluorometric HPLC obtained from HUVECs with or without CRIF1 siRNA. Arrows indicated the signal of total biopterin and BH₂ + B. (C) Quantification of total biopterin, BH₂ + B and BH₄ signal in endothelial cells with or without CRIF1 siRNA (n = 3 per group; ^*P < 0.05 vs. control). (D, E) BH4 synthesis enzymes (GCH-1, PTS, SPR, and DHFR) expression in HUVECs with various amount of CRIF1 siRNA. Densitomeric analysis of each protein was shown in graph (n = 3 per group; ^*P < 0.05, vs. control).