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. 2022 Apr 19;13(4):373. doi: 10.1038/s41419-022-04827-4

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© The Author(s) 2022

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–D Western blot assay confirming the knockdown efficiency of xCT siRNA and xCT shRNA in CRC cells. E–H CCK-8 assay showed that xCT knockdown by both xCT siRNA and xCT shRNA significantly suppressed the proliferation of HCT116 and HCT15 cells. I, J Colony formation assays indicated that xCT inhibition markedly reduced HCT116 and HCT15 cell colony formation ability. K, L EdU assay confirmed that xCT inhibition attenuated HCT116 and HCT15 cell proliferation. M, N Cell cycle assay indicated that xCT inhibition hampered G1/S transition of HCT116 and HCT15 cells. O, P IF staining showed that xCT knockdown diminished the expression of PCNA in HCT116 and HCT15 cells. Q, R Western blot analysis revealed that xCT inhibition downregulated the expression of proliferation-related factors, including PCNA and C-myc, in HCT116 and HCT15 cells. S Western blot analysis indicated that xCT was overexpressed in HCT116 cells upon transfection. T CCK-8 assay showed that overexpression of xCT significantly promoted the proliferation of HCT116 cells. U EdU assay confirmed the promoting effect of xCT overexpression on the proliferation of HCT116 cells. *P < 0.05, **P < 0.01.