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. 2025 Jan 2;44:1. doi: 10.1186/s13046-024-03250-1

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© The Author(s) 2024

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — 130H2 inhibited tumor growth by increasing immune cells infiltration. (A) Experimental schematics of the workflow for in vivo efficacy and TILs analysis. (B and J) BALB/c mice (n = 7) bearing EMT6 cells (B) and CT26 cells (J) were intraperitoneally treated with 10 mg/kg 130H2 or isotype antibody every four days on day 5, 9, 13 post inoculation, three times in total. Significance, *P < 0.05 by Student’s t-test, **P < 0.01 by Student’s t-test. The expression of αvβ8 in the EMT6 cell line and CT26 cell line before inoculation, visualized by staining with 130H2 antibody, is shown in the right. (C-I) BALB/c mice (n = 7 every time in each group) were sacrificed on day 12 and 17 after tumor inoculation and EMT6 tumors were dissociated to analyze tumor-infiltrating immune cells using FACS. Absolute number of cells per gram tumor tissue were calculated for populations of DCs (C) and NK cells (D). Proportion of CD4 + T cell (E) and CD8 + T cell (F) populations were determined by CD3 + CD4 + CD8- and CD3 + CD4-CD8 + markers. Myeloid cell populations including MDSC/myeloid ratio (G), M1/macrophage (H), and M2/macrophage (I) were evaluated. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 were analyzed by Student’s t-test. P < 0.05 means statistical significance. (K) BALB/c mice (n = 6) bearing EMT6 cells were intraperitoneally administrated with the following treatments: 10 mg/kg isotype-hIgG1(day 5, 12) plus 10 mg/kg isotype-rat IgG2a (day 5, 9, 13), 10 mg/kg 130H2 (day 5, 12) plus 10 mg/kg isotype-rat IgG2a (day 5, 9, 13), 10 mg/kg PD-1 antibody (day 5, 9, 13) plus 10 mg/kg isotype-hIgG1 (day 5, 12), 10 mg/kg 130H2 (day 5, 12) plus 10 mg/kg PD-1 antibody (day 5, 9, 13). P < 0.0001 was indicated as ****, P < 0.05 as *, and statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. (L-O) BALB/c mice (n = 6 in each group) were sacrificed on day 15 post-tumor inoculation, and EMT6 tumors were dissociated for analysis of tumor-infiltrating immune cells using FACS. The proportions of CD45 + cell, CD3 + T cell, macrophages, and M1 macrophages were quantified. Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparisons. Data in figures B-O are presented as mean ± SEM