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. 2021 Sep 30;12:5725. doi: 10.1038/s41467-021-25967-9

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© The Author(s) 2021, corrected publication 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. 5 — A Immunofluorescence staining of β1- Adrenergic Receptor (AR), β2-AR, β3-AR, CCL2, CD45, and DAPI in liver tumor tissues from DEN + CCl₄-induced tumor-bearing mice under SE or EE-feeding condition. Original magnification 20 × 10, Scale bar, 100 μm. B Immunohistochemical staining of CCL2 in the tumor tissue from DEN + CCl₄-induced tumor-bearing mice under Standard Environment (SE) and Enriched Enrichment (EE) feeding condition with or without β-AR blockade (β-block, treated with SR59230A + propranolol). The relative CCL2 expression was quantified by Image J analysis (n = 6 for each group). Original magnification 20 ×10, Scale bar, 100 μm. All data are presented as the mean ± SEM, and analyzed by two-tailed unpaired Student’s t test. C Immunohistochemical staining of CCL2 in the tumor tissue from Hepa1-6 tumor-bearing mice under SE- or EE-feeding condition with or without β-AR blockade. The relative CCL2 expression was quantified by Image J analysis (n = 7 for each group). Original magnification 20 × 10, Scale bar, 100 μm. All data are presented as the mean ± SEM, and analyzed by two-tailed unpaired Student’s t test. D Levels of CCL2 in serum from DEN + CCl₄-induced tumor-bearing mice under SE- or EE-feeding condition with or without β-AR blockade (n = 4 for each group). All data are presented as the mean ± SEM, and analyzed by two-tailed unpaired Student’s t test. E ELISA analysis of CCL2 in the liver from DEN + CCl₄-induced tumor-bearing mice under SE or EE-feeding condition with or without β-AR blockade (Vehicle + SE: n = 6, Vehicle+EE: n = 6, β-block+SE: n = 4, β-block+EE: n = 5). All data are presented as the mean ± SEM, and analyzed by two-tailed unpaired Student’s t test. F mRNA expression of CCL2 in the liver from DEN + CCl₄-induced tumor-bearing mice under SE- or EE-feeding condition with or without β-AR blockade (n = 5 for each group). All data are presented as the mean ± SEM, and analyzed by two-tailed unpaired Student’s t test. G–I Mostly tumor cells (CD45^-), immune cells (CD45⁺), TAMs cells (CD45⁺CD11b⁺F4/80⁺Ly6G⁻) and G-MDSCs (CD45⁺ CD11b⁺F4/80⁻Ly6G⁺) in the tumor microenvironment were sorted from subcutaneous Hepa1-6 tumor-bearing mice by flow cytometry after indicated treatments. mRNA expression of CCL2 were determined in tumor cells and immune cells G, TAMs H) and G-MDSCs I. n = 5 for each group. All data are presented as the mean ± SEM, and analyzed by two-tailed unpaired Student’s t test.