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. 2021 Sep 22;40:297. doi: 10.1186/s13046-021-02059-6

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© The Author(s) 2021

Open AccessThis 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. 3 — Knockdown of PTK6 inhibits the stemness of CRC cells. A The GSEA result indicates an enrichment of gene sets related to reactome transportional regulation of pluripotent stem cells in PTK6 overexpression CRC tissues. B The correlations of PTK6 with chemoresistance and stemness related genes were detected from the GEO database. C Kaplan–Meier survival curves in CRC patients undergoing chemotherapy with distinct PTK6 expression levels. D The in vitro limiting dilution assay shows the effects of PTK6 on the formation of CSC spheres (mean ± SD, n = 12), likelihood ratio test. E Tumor sphere formation assays indicate that silencing of PTK6 decreases the sphere formation of CSC cells. Scale bar represents 50 μm. The right panel shows the relative fold change of sphere number (mean ± SD, n = 3). F Western blots assays demonstrate the influence of PTK6 on the expression of stem cell markers in CRC cells. G Real-time q-PCR analysis was used to detect the effect of PTK6 on the expression of stem cell markers and chemoresistance related genes in CRC cells (mean ± SD, n = 3). H Co-immunofluorescent staining of PTK6 (red), CD133 (green) and EPCAM (green) in CRC cells. Scale bar represents 5 μm. I Co-immunofluorescent staining of PTK6 (red), CD133 (green) and EPACAM (green) in tumor spheres. Scale bar represents 5 μm. J The number of CD133⁺ or SOX2⁺ cells was evaluated in the control and PTK6 silencing CRC cells by flow cytometry (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001