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. 2021 May 5;12:2537. doi: 10.1038/s41467-021-22755-3

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© The Author(s) 2021

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 — a Overall procedures to establish 4T1-BALB/c orthotopic metastasis model. Cancer cell inoculation was followed by treatment with YB1 or PBS. b Quantification of 4T1 primary tumor size (two-sided multiple t-tests) and relevant lung metastasis (unpaired two-tailed t-test) after YB1 treatment in the 4T1-BALB/c orthotopic metastasis model (n = 6 PBS, n = 7 YB1). c Mice tumor size measured (unpaired two-tailed t-test) before surgical removal of 4T1 primary tumors and relevant Kaplan–Meier survival curve (log-rank test) of these mice (n = 8 PBS, n = 9 YB1). d Procedure to establish 4T1-BALB/c experimental metastasis model. 4T1 cells were i.v. injected into BALB/c mice pretreated with YB1 or PBS. e Picture of lungs fixed in Bouin solution and quantification (unpaired two-tailed t-test, displayed is a combined result of three independent experiments, n = 16 per group) of 4T1 lung metastases after YB1 treatment in the 4T1-BALB/c experimental metastasis model. f Picture of lungs and quantification (unpaired two-tailed t-test, n = 11 per group) of lung metastases after YB1 treatment in an experimental metastasis model established by colon cancer (CT26) in BALB/c mice. g Picture of 4% PFA-fixed lungs and representative H&E staining of lung tissue in an experimental metastasis model established by bladder cancer (MB49) in C57BL/6J mice (n = 4 mice per group). Scale bar, 3 mm. Condensed tumor nodules were stained with H&E from PBS samples. h Picture of lungs and quantification (unpaired two-tailed t-tests) of lung metastases after YB1 treatment in the experimental metastasis model established by inoculation of melanoma (B16F10) in C57BL/6J mice (n = 8 PBS, n = 9 YB1). i Quantification of 4T1 lung metastases in the 4T1-BALB/c experimental metastasis model after treatment with different doses of YB1 (n = 5 per group). P-values are comparisons between no YB1 treatment and different dosages of YB1 treatment using unpaired two-tailed t-tests. Graphs and images presented are one representative experiment of two independent experiments (b, c, g, i). Graph presented is a combined result of two independent experiments (f, h). All data are presented as mean values ± SEM. Source data are provided as a Source Data file.