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. 2024 Jan 9;15(1):24. doi: 10.1038/s41419-023-06408-5

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© The Author(s) 2024

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. 3 — A HCT15 and HCT116 cells were treated with TNFα (50 ng/mL) and OXP or 5-Fu for 24 h. Cell viability was evaluated by CCK assay (n = 3). ***p < 0.001. Two-Way ANOVA test. B HCT15 and HCT116 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 24 h. Caspase-3 activity was evaluated with a caspase-3 activity kit (n = 3). *p < 0.05 and **p < 0.01. One-Way ANOVA test. C HCT15 and HCT116 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 24 h. The cleavage of caspase 8, caspase-3 and PANX1 was evaluated by western blotting (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001. One-Way ANOVA test. D After treatment with TNFα and OXP for 6 h, the intracellular ATP content was examined by flow cytometry (n = 3). *p < 0.05. One-Way ANOVA test. E HCT15^shNC and HCT15^shPANX1 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 6 h. The intracellular ATP content was examined by flow cytometry (n = 3). *p < 0.05. One-Way ANOVA test. Similar experiments were carried out in HCT116 cells. F HCT15^shNC and HCT15^shPANX1 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 6 h. The extracellular ATP content was examined with a luminescent ATP detection kit (n = 3). *p < 0.05 and ***p < 0.001. One-Way ANOVA test. Similar experiments were carried out in HCT116 cells. G Schematic diagram of the DC maturation analysis. HCT116^shNC and HCT116^shPANX1 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 24 h. Then, these cells were cocultured with THP1- iDCs for 24 h. H The mRNA levels of the DC maturation marker CD86 and the proinflammatory cytokines IL-6, IL-1β and IL-18 in THP-iDCs that were cocultured with vehicle- or OXP/TNFα-treated HCT116^shNC and HCT116^shPANX1 cells were evaluated by qRT‒PCR (n = 3). ***p < 0.001. One-Way ANOVA test. I HCT116^shNC and HCT116^shPANX1 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 24 h. Then, these cells were cocultured with THP1-iDCs for 24 h and Jurkat T cells for 15 h. J The mRNA levels of the cytotoxic cytokines IFNγ and GzmB in Jurkat T cells that were cocultured with THP-iDCs and vehicle- or OXP/TNFα-treated HCT116^shNC/HCT116^shPANX1 cells was evaluated by qRT‒PCR (n = 3). ***p < 0.001. One-Way ANOVA test.