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. 2023 Jun 12;30:39. doi: 10.1186/s12929-023-00937-z

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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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — KLF10 competed with ELF3 and transcriptionally suppressed Notch-3 and -4. A Primer maps with predicted SP-1/KLF10 binding sites (left panel, predicted p < 0.001) and various designed deletion (right panel) of promoter regions of Notch-3 and -4 for luciferase reporter assay. B Luciferase activity of Panc-1 cells transfected with vector control (yellow), Notch-3(blue), and Notch-4(green) gene promoter plasmids without (filled) or with (diagonal stripe) hemagglutinin (HA) tagged-KLF10 transfection (OE). The bar graphs were mean ± SE from cumulated data of three independent experiments. *p < 0.05 and **p < 0.01, respectively. C Luciferase activity of Panc-1 cells, transfected with Notch-3 (yellow) or Notch-4 (blue) gene promoter plasmids with full length (filled) or with various deletion (diagonal stripe, diagonal check) indicated. The bar graphs were mean ± SE from cumulated data of three independent experiments. *p < 0.05 and **p < 0.01, respectively. D (Upper panel) primer maps of Notch-3/4 gene promoters for CHIP-PCR assay. (Lower panel) CHIP assay. DNA fragments of Panc-1 cells were immune-precipitated with KLF10, IgG or positive control (P) followed by PCR amplification of the Notch-3/4 gene promoter region that contains KLF10 binding sites (input), as described in “Materials and methods” section. E Quantitative PCR of the Notch-3/4 gene promoter region using samples from MiaPaCa (yellow) or Panc-1 (blue) cells prepared as described in D. The bar graphs were mean ± SE from cumulated data of three independent experiments. F MiaPaCa cells with conditional over-expression of KLF10 under Dox treatment were immuno-precipitated with KLF10 antibody and quantitative PCR of the Notch-3 (yellow) and Notch-4 (blue) gene promoter regions. The bar graphs were mean ± SE from cumulated data of three independent experiments. *p < 0.05. G Primer maps of predicted ELF3 binding sites on the promoter regions of Notch-3 (upper panel) and Notch-4 (left panel) genes. H Luciferase reporter assays of Notch-3 (left panel) and Notch-4 (right panel) under conditions of various combination of HA-KLF10 and ELF3 (yellow) and ELF3-mutant (ELF3-S68A, blue) as indicated. The bar graphs were mean ± SE from cumulated data of three independent experiments. **p < 0.01