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. 2023 Sep 22;101(11):1449–1464. doi: 10.1007/s00109-023-02369-6

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — NF-kB directly regulates LOXL2 expression or indirectly through ZEB1. A Immunoblotting for phospho NF-κB and NF-κB using nuclear and cytoplasmic fractions prepared from Mia GR cells after treatment with JSH-23 for 4 h. Relative mRNA expression (B), and protein expression (C), of LOXL2 and ZEB1 in Mia Con, Mia GR (no treatment, JSH-23 1 h or 4 h treatment) cells. D Scheme for the expected NF-kB binding site on the LOXL2 gene promoter. E ChIP fold enrichment of LOXL2 DNA fragments (NP1, NP2, NP3) by chIP-qPCR. F Changes in chIP fold enrichment with NF-kB in the NP2 region on the LOXL2 gene promoter in Mia Con and Mia GR cells (no treatment, 2DG treatment, and siGLUT3 transfection) (G), Mia GR cell transfected with LOXL2 and ZEB1 siRNA. Knockdown efficiencies were evaluated by immunoblotting using LOXL2 and ZEB1 antibodies. H Relative gene expression of LOXL2 and ZEB1 confirmed by qPCR in Mia GR transfected with siRNA. I Scheme for the expected ZEB1 binding site on the LOXL2 gene promoter. J ChIP fold enrichment of LOXL2 DNA fragments (ZP1, ZP2, ZP3) by chIP-qPCR. K Changes in chIP fold enrichment with NF-kB in the ZP3 region on the LOXL2 gene promoter in Mia Con and Mia GR cells (no treatment, JSH-23 4 h treatment)