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. 2020 Mar 13;11:1364. doi: 10.1038/s41467-020-15186-z

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© The Author(s) 2020

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. 3 — a Representative image of G-CSF mRNA expression in ME180-Control or ME180-GCSF cells as evaluated by RT-PCR. b Representative G-CSF staining in ME180-Control cells or ME180-GCSF cells-derived tumors. c WBC/granulocyte counts of ME180-Control-derived tumor-bearing rats and ME180-GCSF-derived tumor-bearing rats (three rats per group). Rats were subcutaneously inoculated with ME180-Control or ME180-GCSF cells. Three weeks after inoculation, their subcutaneous tumors or peripheral blood samples were collected for analyses. d 18F-FDG-PET/CT scan of ME180-Control-derived tumor- and ME180-GCSF-derived tumor-bearing rats. Rats were subcutaneously inoculated with ME180-Control or ME180-GCSF cells. Three weeks after inoculation, 18F-FDG-PET/CT was performed. ME180-GCSF-derived tumor-bearing rats showed significant 18F-FDG-uptake in the PALN (circle). e PALNs resected after 18F-FDG-PET/CT. f Representative pathological findings from the PALNs resected after 18F-FDG-PET/CT (hematoxylin and eosin). Both of the images contain no malignant cells. g Effects of tumor-derived G-CSF on the induction of MDSC in rat models of cervical cancer. CD11b/c⁺HIS48⁺ cell populations detected in the peripheral blood and lymph nodes. Rats were subcutaneously inoculated with ME180-Control or ME180-GCSF cells. Three weeks after inoculation, the number of MDSC was evaluated by flow cytometry (three rats per group). h Effects of tumor-derived G-CSF and anti-Gr-1 antibody on PALN in mice models of cervical cancer. i Effects of anti-Gr-1 antibody on MDSC induction in mice models of cervical cancer. CD11b⁺Gr1⁺ cell populations detected in the peripheral blood and lymph nodes. h, i ME180-Control- or ME180-GCSF-derived tumor-bearing mice were treated with either control IgG or anti-Gr-1 antibody for three weeks (five mice per group). Then, the number of MDSC was evaluated by flow cytometry (five mice per group). j Ability of CD11b⁺Gr-1⁺ cells to suppress CD8⁺ T cell assessed by T-cell proliferation assay. CD11b⁺ Gr-1⁺ cells (MDSC) were isolated from spleen of ME180-GCSF-derived tumor-bearing mouse. CD8⁺ T cells (2 × 10⁵ cells/well) were isolated from spleen of Balb/c mice and co-cultured with MDSC at indicated ratios. Cells were incubated for 72 h, after which BrdU was added for an additional 24 h. T cell proliferation was determined by BrdU incorporation (n = 6). Error bars indicate mean ± SD. Statistical significance was assessed using two-sided Welch t test. bp, base pairs. Scale bar, 50 μm. Source data are provided as a Source Data file.