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. 2023 May 19;14:2861. doi: 10.1038/s41467-023-38581-8

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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 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. 1 — GC patient-derived organoids of intestinal subtype GX006, GX055 and GX060 were trained with increasing concentrations of 5-fluorouracil (5FU) and cisplatin (CDDP) combination to develop 5FU + CDDP resistant lines. a CellTiter-Glo analysis showing the viability of parental versus 5FU + CDDP resistant organoids following treatment in various concentrations of 5FU + CDDP combinations. b Annexin V-PI analysis showing the percentage of apoptotic cells in parental versus 5FU + CDDP resistant organoids following treatment in 1.25 µM 5FU + 5 µM CDDP (GX006) or 5 µM 5FU + 20 µM CDDP (GX055 and GX060). c In vitro limiting dilution spheroid formation and tumor-initiating cell frequency calculation in parental versus 5FU + CDDP resistant organoids. d, e Gene Ontology (GO) (d) and Gene Set Enrichment Analysis (GSEA) (e) of differentially expressed genes identified by RNA-seq data found enrichment of interferon signaling and its downstream JAK/STAT signaling in the 5FU + CDDP resistant organoids as compared to parental controls. f Western blot for phosphorylated and total JAK2, phosphorylated and total STAT3, ADAR1 and ADAR2 in the three paired parental and 5FU + CDDP resistant organoid lines. β-actin served as a loading control. g Western blot for phosphorylated and total JAK2, phosphorylated and total STAT3 and ADAR1 in the three parental organoid lines with or without interferon (1000 U/mL) treatment for 24 hours. h Western blot for total STAT3 and ADAR1 in the GX006 parental and GX006 5FU + CDDP resistant organoid lines stably transduced with non-target control (NTC) or STAT3 shRNA knockdown (clones 1 and 2) after treatment with vehicle control (CTRL) or 1000 U/mL interferon (IFN) for 24 hours. β-actin served as a loading control. Images representative of n = 3 independent experiments. (a) n = 2 independent experiments; (b) n = 3 independent experiments for GX006, GX005 and n = 5 independent experiments for GX060; (c, f, g, h) n = 3 independent experiments. Significance were calculated by (b) unpaired two-tailed student t-test; (c) one-sided extreme limiting dilution analysis. Data was presented as mean ± standard deviation. NES for normalized enrichment score, FDR for false discover rate. Source data are provided as a Source Data file.