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. 2022 Apr 6;29(10):1439–1451. doi: 10.1038/s41417-022-00453-6

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© The Author(s) 2022

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. 5 — A BALB/C nude mice were pretreated with exosomes derived from the indicated cells every two days for four times, and then the collected lung and liver tissues were subjected to double-label immunofluorescence for VE-cadherin (green) and CD31 (red). Structures positive for CD31 are indicated by arrowheads. The scale bar represents 100 μm. B The vasculature exhibited higher permeability after treatment by HMGA2-overexpressing exosomes. Mice pretreated with exosomes with high or low HMGA2 expression were intravenously injected with rhodamine-dextran, and frozen tumor tissue sections were used for imaging by fluorescence microscopy. Rhodamine-dextran exhibited red fluorescence and cell nuclei were stained blue with DAPI. Scale bar, 50 μm. C Gross appearance of the liver and lung in the tail vein injection metastatic model and metastatic lesion quantitative analysis. Data are presented as the mean ± SD (n = 6 biologically independent animals). ***p < 0.001. D Representative H&E-stained metastatic tumor nodules in the liver and lung and quantitative analysis of metastatic nodules. Data are presented as mean ± SD (n = 6 biologically independent animals). ***p < 0.001. E The expression of HMGA2 in mouse serum-derived exosomes from mice bearing tumors derived from 5-8F cells with and without HMGA2 overexpression. F The tumor mass of subcutaneous xenograft tumors derived from of 5-8F cells with and without HMGA2 overexpression was transplanted into the liver of BALB/C nude mice. Collected lungs were subjected to double-label immunofluorescence for ZO-1 (green) and CD31 (red). Structures positive for CD31 are indicated by arrowheads. The scale bar represents 100 μm. G The lung vasculature permeability in BALB/C nude mice bearing 5-8F HMGA2 OE control or 5-8F HMGA2 OE tumors was measured by rhodamine-dextran in lung tissue. Rhodamine-dextran exhibited red fluorescence and cell nuclei were stained blue with DAPI. Scale bar, 50 μm. H 5-8F/vec or 5-8F/HMGA2 tumors were injected into the liver, and gross appearance of the liver and liver weight quantitative analysis in different groups are shown. I Gross appearance of the lung in the liver metastatic model and metastatic lesion quantitative analysis. Data are presented as the mean ± SD (n = 6 biologically independent animals). ***p < 0.001. J Quantitative analysis of representative H&E-stained the number of metastatic tumor nodules in the lung and metastatic nodules. Data are presented as the mean ± SD (n = 6 biologically independent animals). ***p < 0.001.