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. 2019 Jun 28;47(16):8424–8438. doi: 10.1093/nar/gkz560

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© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — NF-κB response elements mediate the expression of ENPP2. (A) Luciferase assay demonstrating increased transcriptional activity at the ENPP2 promoter following LPS treatment for the indicated time. (B and C) Endogenous ENPP2 expression in HEK 293T cells treated with LPS at the indicated time points (B) qPCR analysis (C) representative western blots and quantifications. (D) Effect of a 24 h treatment with BAY11-7085 on LPS-mediated ENPP2 expression, qPCR analysis. (E) Effect of p65 overexpression on ENPP2 promoter activity as shown by luciferase reporter assay. (F) Expression of endogenous ENPP2 following p65 overexpression in HEK 293T, qPCR analysis. (G) Scheme of the two кB consensus sites localization in the ENPP2 promoter. (H) ChIP-qPCR measurement of p65 enrichment at the ENPP2 locus in HEK 293T cells treated with LPS for 6 h. A set of primer targeting an intergenic region downstream of SNRPN was used a negative control (NEG). (I) Scheme of the ENPP2 luciferase reporter. (J) Luciferase reporter assay showing the effect of the кB sites deletion on ENPP2 promoter activity. LPS 100ng/ml, BAY11-7085 1 μM.