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. 2023 Nov 29;14:7863. doi: 10.1038/s41467-023-43670-9

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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. 2 — a Experimental scheme: differential expressed genes (DEGs) analysis in Prom1+ tumor-initiating/propagating cells using two HCC mouse models (DEN+CCl₄-induced fibrosis-related HCC and NRasV12+Myr-AKT HTVI) and one liver regeneration model (0.1% DDC diet). Transcriptome sequencing identified Prom1+ cells-specific DEGs in HCC tumors, but not in regenerating liver. TCGA-LIHC cohort used for expression and clinical survival analysis to narrow down potential candidate genes. See Supplementary Fig. S2A for detailed data analysis pipelines. SB = Sleeping beauty b qPCR validation for Spink1 expression in the CD133+ and CD133- subpopulations in the three mouse models. c qPCR analysis for Spink1 expression in non-tumor (NT) and tumor tissues (T) of the two HCC mouse models, and in sham or regenerating livers of 70% partial hepatectomy (PH) mouse model. d RNA-seq analysis for Spink1 mRNA expression in various tissues of nonalcoholic steatohepatitis (NASH) mouse model. e, f Pearson correlation analysis of Spink1 and Prom1 expression in two HCC models, from normal to advanced HCC (●, ■, ▲, ▼, ○, □, △, ▽). g, h qPCR analysis for Spink1 expression in Ctrl and Prom1-DTA mice of the two mouse models. i Representative liver tissue images from Ctrl or Prom1-DTA mice of NRasV12+Myr-AKT mouse model treated with DMSO or 5-FU, showing Spink1 (red) and CD133 (green). DAPI (blue), nucleus. Scale bar in low magnification = 30 μm; high magnification = 2 μm. b n = 2 mice; c n = 3 mice (T and NT) of DEN+CCl₄ model and liver regeneration model, n = 3 mice (NT), 6 mice (T) of NRasV12+Myr AKT model; d n = 5 mice; e, f n = 2–7 mice, g, h n = 3 mice (Ctrl), 4 mice (Prom1-DTA) of DEN+CCl₄ HCC model, n = 4 mice (Ctrl), 3 mice (Prom1-DTA) of NRasV12+Myr AKT HCC model. Data were expressed as mean ± s.e.m. Statistical analysis: b, c two-way ANOVA, d one-way ANOVA, e, f two-sided Pearson correlation analysis, or g, h two-sided Unpaired Student’s t-test. Source data are provided as a Source Data file. Illustration for a was created using BioRender.com.