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. 2022 Jun 27;13:3671. doi: 10.1038/s41467-022-31238-y

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© The Author(s) 2022, corrected publication 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. 4 — a Representative histogram of cell surface MHC-I detected by flow cytometry. Adjacent bar graph displaying geometric mean fluorescence intensity (GMFI). Line represents median, data points represent individual animals, saline (n = 5), CpG (n = 6), Mann–Whitney test, exact p-value. b Survival (time to endpoint length: 20 mm) for animals given CpG/anti-OX40 (aOX40) (n = 10) or isotype/vehicle (n = 9). Log rank test, exact p-value. c Tumor volume following treatments (Rx) indicated by arrows. Isotype (n = 6), CpG (n = 5), aOX40 (n = 6), CpG/aOX40 (n = 5). Mean ± S.E.M, two-sided, unpaired t-test comparing tumor volume on day 10 in isotype treated vs. CpG/aOX40, exact p-value. d–g Flow cytometry analysis of immune cells found in the tumor post-treatment initiation day 10. Isotype (n = 6), CpG (n = 5), anti-OX40 (n = 6), CpG/aOX40 (n = 8). Mean ± S.E.M., Mann–Whitney test., exact p-values: d Percent of live cells in the tumor that are T-cells. e Percent of CD4+ (gating: Singlets/Live/TCRβ+, CD45+/CD8−, CD4+) T-cells that are CD25+ and FOXP3+. f Ratio of CD8+ (gating: Singlets/Live/TCRβ+, CD45+/CD8+, CD4−) T-cell counts to Treg counts (gating: Singlets/Live/TCRβ+, CD45+/CD8−, CD4+/CD25+, FOXP3+). g Geometric mean fluorescence intensity for intracellular granzyme B in the CD8+ T-cells (gating: Singlets/Live/TCRβ+, CD45+/CD8+, CD4−). h MTB/TOM tumor volumes graphed to endpoint: isotype (n = 6), anti-PD-L1 (n = 7), CpG/aOX40 (n = 6), and triple combination (n = 9). i Tumor volume in MTB/TOM animals after last day of treatment (related to panel h). Mean ± S.E.M., two-sided, unpaired t-test, exact p-value. j–l Survival (time to ethical endpoint length = 20 mm). Log rank test for each treatment arm in comparison to isotype arm, exact p-value in black. Log rank test for anti-PD-L1 compared to triple combination, exact p-value in red: j MTB/TOM (MYC-ON) animals from panel h. k MC38-MYC model, isotype (n = 12), anti-PD-L1 (n = 11), CpG/aOX40 (n = 10), triple combination (n = 11). l WAP-MYC model, isotope (n = 9), anti-PD-L1 (n = 6), CpG/anti-OX40 (n = 9), and triple combination (n = 8). m Disease-free survival in MTB/TOM model (defined as time to palpable tumor). Mice previously treated with triple combination and eradicated their tumors were transplanted with a new MTB/TOM tumor on the contralateral side. Tumor naïve group (n = 5) and responded to therapy group (n = 5). Log rank test, exact p-value. Source data are provided in the Source Data file.