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. 2024 Jan 10;56(1):192–209. doi: 10.1038/s12276-023-01147-1

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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 Establishment of TRAIL-resistant H292 (H292R) and HepG2 (HepG2R) cells and identification of TRAIL sensitivity. Cell viability was assessed by MTT assay. Parent H292 (H292S) and H292R cells or parent HepG2 (HepG2S) and HepG2R cells were treated with TRAIL or TNF-α at the indicated concentrations for 24 h. Data are shown as the means ± SDs of three independent assays. The statistical significance of differences between two groups was determined with the two-tailed Student’s t test. b TNFSF-mediated tumor cell killing assay in H292S and H292R cells or HepG2S and HepG2R cells. Cells were exposed to TRAIL or TNF-α at the indicated concentrations for 24 h, and cytotoxicity was analyzed by propidium iodide and Annexin V staining assays. c Immunoblot analysis of TNFSF-mediated activation of caspases in H292S and H292R cells or HepG2S and HepG2R cells. Cells were treated with TRAIL or TNF-α at the indicated concentrations for 24 h, and the activation of caspases in cell lysates was analyzed by immunoblotting. d Analysis of PRKCSH mRNA and protein expression in H292S and H292R cells or HepG2S and HepG2R cells. Data are representative immunoblots and quantitative analysis of PRKCSH expression levels in each cell line. e TNFSF-mediated tumor cell killing assay in H292R or HepG2R cells. Cells transfected with siPRK#2 or siCon were treated with TRAIL or TNF-α at the indicated concentrations for 24 h. TNFSF-mediated cytotoxicity against tumor cells was analyzed by MTT assay (for cell viability) and propidium iodide and Annexin V staining assay (for cell death). f Immunoblot analysis of TNFSF-mediated activation of caspases in H292R or HepG2R cells. Cells transfected with siPRK#2 or siCon were treated with TRAIL or TNF-α at the indicated concentrations for 24 h, and the activation of caspases in cell lysates was analyzed by immunoblotting. **p < 0.01, *p < 0.05.