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. 2023 Sep 15;9:95. doi: 10.1038/s41421-023-00592-6

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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 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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Fig. 5 — a Top: the strategy to construct truncated N1ICD plasmids; Bottom: co-IP followed by western blotting assay to detect N1DARP-interacting N1ICD truncations. b Notch signaling and its crosstalk with other pathways detected using western blotting assay using Capan1 transfected with N1DARP and individual domain of N1ICD. c Molecular docking using CABS-dock and PyMOL assessing electrostatic attraction and identifying key amino acids that mediate N1DARP–ANK interaction. d Western blotting assay detecting N1DARP–N1ICD interaction in HEK293T transfected with wild-type N1DARP or its key amino acid mutants. e Stem cell properties assessed by western blotting assay using Capan1 with wild-type N1DARP or its Y20/Y34 mutant. f The segregating effect of N1DARP on RBPJ–N1ICD interaction detected using western blotting assay. g N1DARP–N1ICD interaction detected using western blotting assay with Capan1 transfected with wild-type N1ICD or its M1 and M2 mutants. h Notch signaling and its crosstalk with other pathways detected by western blotting assay using Capan1 transfected with N1DARP or N1ICD-M1M2 mutant. i, j Cell proliferation detected using (i) colony formation assay and (j) chemosensitivity assessed using IC₅₀ assay using Capan1 transfected with N1DARP or wild-type N1ICD and its M1M2 mutant. k The tumor volume of subcutaneously injected xenograft model using Capan1 transfected with N1DARP or N1ICD wild-type and its M1M2 mutant. l The apoptosis rate detected by flow cytometry using subcutaneously injected tumor derived from Capan1 transfected with N1DARP or wild-type N1ICD and its M1M2 mutant on day 35 after transplantation. m Notch signaling and its crosstalk with other pathways detected by western blotting assay using N1ICD-knockout Capan1 transfected with N1DARP and wild-type N1ICD or its M1M2 mutant. The data are presented as the mean ± SD of three independent experiments. ns no significance; *P < 0.05, **P < 0.01 by Student’s t-test (i, l); *P < 0.05, **P < 0.01 by one-way ANOVA (j, k).