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. 2023 Oct 23;14:6690. doi: 10.1038/s41467-023-42332-0

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — A Representative H&E staining of metastatic livers from C57BL/6 J mice intraportally implanted with the control (Ctrl), OTUD1 knockdown, FGL1 knockdown or FGL1 knockdown with overexpression of short hairpin RNA (RNA)-resistant FGL1 (rFGL1) MC38 cells followed by TNFα treatment. Scale bar for bright-field images, 1 cm; scale bar for H&E images, 2.5 mm. Right panel, experimental strategies. B Quantification of liver weight and the number of liver metastases of the indicated groups of mice (n = 5 mice per group). C Flow cytometric analysis of the number of IFN-γ⁺ CD8⁺, IFN-γ⁺ CD4⁺ and Ki67⁺ CD8⁺ T cells in liver metastases of indicated groups of mice (n = 5 mice per group). D Survival curve analysis of mice in indicated groups (Mean survival times of Ctrl+PBS, Ctrl+TNFα, shOTUD1+TNFα, shFGL1+TNFα and shFGL1 + rFGL1+TNFα mice were 25.8, 19.1, 27.6, 32.5 and 21.5 days, respectively) (n = 8 mice per group). E Representative H&E staining of metastatic livers from C57BL/6 J mice intraportally injected with MC38 cells followed by treatment with the control (IgG), anti-CSF1R and/or anti-PD-1 monoclonal antibodies. Scale bar for bright-field images, 1 cm; scale bar for H&E images, 2.5 mm. Right panel, experimental strategies. F Quantification of liver weight and the number of liver metastases of indicated groups of mice (n = 5 mice per group). G Representative H&E and IHC staining of the indicated markers in CRC metastatic liver tissues from patients with high (N = 14) or low (N = 13) plasma FGL1 levels detected in Fig. 2 (C) (N = 27). Scale bar, 50 μm. H Percentage of specimens with high or low CD68, P-p65, OTUD1, CD4, and CD8 expression in the low or high plasma FGL1 groups. I OTUD1 expression is inversely correlated with overall survival in patients treated with anti-PD-L1 in the study by Snyder and colleagues on urothelial cancer (UC_Synder, N = 25). The data in (B, C, F) are presented as the mean ± SD. The data in (D, I) were determined by Kaplan‒Meier analysis with the log-rank test. The data in (H) were determined by the two-sided chi-square test. P values were determined by one-way ANOVA (B, C, F).