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. 2024 Feb 24;15:1700. doi: 10.1038/s41467-024-45996-4

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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/.

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Fig. 7 — a WT or IFNAR1 KO mice were implanted with MC38 tumor cells s.c. and treated with 25 mg/kg b.i.d. ceralasertib. Splenocytes were restimulated with tumor-specific (p15e) or irrelevant peptides (mGp100). Number of IFN-γ spots are shown. N = 12 WT, N = 6 IFNAR1 KO. Individual mouse results, mean and SEM are shown. Two-sided unpaired Student’s t-test was used. NS—non-significant (p > 0.05). b T-cells isolated from LN of mice treated with 10 mg/kg anti-PD-L1, 25 mg/kg b.i.d. ceralasertib, or combination were plated in the presence of feeder of control splenocytes, restimulated for 6 h with cell-stimulation cocktail, and stained for intracellular IFN-γ. Fold increase of % of IFN-γ⁺ cells within CD8⁺ T-cells is shown. N = 5. Individual mouse results, mean and SEM are shown. One-way ANOVA with correction for multiple comparisons was used. Non-significant p values (p > 0.05) are not shown. c DC were isolated from LN of MC38-bearing mice (C57BL/6) treated with 25 mg/kg b.id. ceralasertib and plated with BALB/c derived splenocytes labeled with Cell Trace Far-Red at a 1:5 (DC:splenosytes) ratio. After 5 days proliferation of CD8⁺ and CD4⁺ T-cells was measured by Cell-Trace dilution. N = 4. Individual mouse results, mean and SEM are shown. Two-sided unpaired Student’s t-test was usede. d Experiments were performed as in (c) except DC were isolated from LN of IFNAR1 KO MC38-bearing mice. N = 3—vehicle group, N = 4—Cerala group, N = 6—no DC group. Individual mouse results, mean and SEM are shown. e PMN-MDSC were isolated from tumors of mice treated for 7 days with 25 mg/kg b.i.d. ceralasertib and plated at different ratios with PMEL splenocytes in the presence of specific peptide. After 48 h the number of CD8⁺ T-cells was calculated by flow cytometry. N = 10. Individual mouse results, mean and SEM are shown. Two-sided unpaired Student’s t-test was used. Non-significant (p > 0.05) p values are not shown. f IFNI signature heatmap evaluated in PMN from naïve mice (N = 3), tumor PMN-MDSC isolated from mice treated with vehicle (N = 3) or with 25 mg/kg ceralsertib for 7 days (N = 3). Source data are provided as a Source Data file.