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. 2023 Mar 31;14(3):229. doi: 10.1038/s41419-023-05756-6

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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. 2 — A Western blotting was performed to examine the protein expression of Nodal in Nodal-overexpressing HCT116 cells and SW480 cells (vector group: cells transfected with an empty vector; oe-Nodal group: cells transfected with the Nodal lentiviral overexpression vector). B Representative images of Nodal expression detected in Nodal-overexpressing HCT116 cells and SW480 cells via IF staining. C, D CCK-8 and colony formation assays validated the increased proliferative ability of HCT116 and SW480 cells after Nodal overexpression. E Representative images for the xenograft tumours morphology. F, G The volume and weight of tumours were evaluated in two groups. H Western blotting was performed to examine the expression of Nodal, p-Smad2/3 and Smad2/3 in each xenograft tumor tissue. I The Nodal, Ki-67, E-cadherin, p-Smad2/3, and Smad2/3 expressions in xenograft tumour tissues were detected via IHC staining. J Wound healing assay revealed that Nodal overexpression increased the migration ability of HCT116 and SW480 cells. K Transwell assay validated the increased migration and invasion ability of HCT116 and SW480 cells after Nodal overexpression. L Western blotting was performed to examine the E-cadherin, N-cadherin and Snail expressions of HCT116 and SW480 cells in the vector/oe-Nodal group. M Representative fluorescence images of lung metastasis in mice and the relative luminescence intensity. (^∗∗P < 0.01 and ^∗∗∗P < 0.001).