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. 2023 Oct 23;14:6699. doi: 10.1038/s41467-023-42360-w

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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. 2 — a The CDK6 protein levels in the non-target control (NTC) and shCDK6 (#1 and #2) MHCC-97L and Hep3B cells as well as in the control (sgCTRL) and sgCDK6 (#1 and #2) subclones derived from PLC/PRF/5 cells were determined by western blot analysis (n = 3 independent experiments). b In vitro limiting dilution sphere analysis showed the role of CDK6 in the regulation of self-renewal ability (n = 2 independent experiments, one-sided extreme limiting dilution analysis). c Left and middle: Knockdown of CDK6 in MHCC-97L and Hep3B cell lines suppressed tumorigenicity compared to NTC cells. Right: Overexpression of CDK6 led to increased tumorigenicity of PLC/PRF/5 cells. Scale bar = 1 cm. d The expression of liver CSC markers, including CD47 (MHCC-97L: n = 4; Hep3B: n = 6 and PLC/PRF/5: n = 3 independent experiments) and CD90 (MHCC-97L: n = 5; Hep3B: n = 7 and PLC/PRF/5: n = 3 independent experiments), was measured by flow cytometry analysis (two-tailed t test). e The migration and invasive abilities of HCC cells were evaluated by uncoated (top; MHCC-97L and PLC/PRF/5: n = 3 and Hep3B: n = 6 independent experiments) and Matrigel-coated transwell (bottom; MHCC-97L: n = 4; Hep3B: n = 7 and PLC/PRF/5: n = 3 independent experiments) assays, respectively. Scale bar = 250 µm. (two-tailed t test). Data was presented as mean ± standard deviation. Source data are provided as a Source Data file.