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. 2023 Aug 29;20(1):114–130. doi: 10.1080/15548627.2023.2249762

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© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 2. — Effects of histone lactylation inhibition on colorectal cancer cells proliferation and survival in hypoxia.

Note: (A) Western blotting analysis of lactylation or H3K18la levels in normal colon epithelial cells and CRC cells. (B) Western blotting analysis of Pan- and site-specific histone lactylation in HCT116 and SW620 cells in normoxia compared with in hypoxia (1% oxygen, 24 h). (C) Lactylation and H3K18la levels were detected in HCT116 and SW620 cells cultured in different concentrations of 2-DG or oxamate for 24 h in hypoxia (1% oxygen) by western blot. (D) Lactylation and H3K18la levels were detected in HCT116 and SW620 cells cultured in hypoxia (1% oxygen) at the indicated time. (E and F) Proliferation of HCT116 and SW620 cells cultured in different concentrations of 2-DG (E) or oxamate (F) in hypoxia (1% oxygen) was analyzed using CCK8 assay. ***P ≤0.001. (G) Tumor growth of HCT116 and SW620 cells treated with different concentrations of 2-DG or oxamate in hypoxia (1% oxygen) was evaluated by colony formation assay. (H) Statistical analysis of the colony formation assay performed using HCT116 and SW620 cells treated with different concentrations of 2-DG or oxamate in hypoxia (1% oxygen). All of the experiments were performed in triplicate, and relative colony numbers are shown as means ± SD. ***P ≤0.001. (I) ANXA5/annexin V-FITC and PI staining showing apoptosis in HCT116 and SW620 cells cultured in different concentrations of 2-DG or oxamate in hypoxia (1% oxygen). (J) Quantification of apoptotic cells. ***P ≤0.001. (K) Western blotting analysis of cleaved CASP3 and cleaved PARP in in HCT116 and SW620 cells cultured in different concentrations of 2-DG or oxamate in hypoxia (1% oxygen).