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. 2020 Mar 12;11:1335. doi: 10.1038/s41467-020-14987-6

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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. 4 — a Survival curves of RT2 or RT2.Stat1^−/− mice treated with either isotype control mAbs (Ctr, RT2 N = 9, RT2.Stat1^−/− N = 4), with adoptive transfer of T antigen-specific CD4⁺ T_H1 cells (AT, RT2 N = 15, RT2.Stat1^−/− N = 9), with immune checkpoint inhibitors (ICB, RT2 N = 12, RT2.Stat1^−/− N = 4; anti-PD-L1 and anti-LAG-3) or with ICB/AT (RT2 N = 15, RT2.Stat1^−/− N = 10). For each AT experiment we generated a non-cytotoxic Tag-specific T_H1 cell line. b Cancer size shown by representative hematoxylin and eosin staining of pancreas of RT2 (upper panel) or RT2.Stat1^−/− mice (lower panel) after four weeks of treatment. Mice were treated as described in a. Green lines depict the size of RT2-cancers after treatment. After 4 weeks of treatment ICB/AT-treated mice (N = 6) had a two-fold smaller islet size than Ctr (N = 7). c Time course of blood glucose levels as surrogate marker for the growth of the insulin-producing tumours of RT2 or RT2.Stat1^−/− mice (median ± interquartile range) treated as described in (a Ctr, RT2 N = 7, RT2.Stat1^−/− N = 4; AT, RT2 N = 10, RT2.Stat1^−/− N = 9; ICB, RT2 N = 6, RT2.Stat1^−/− N = 4, ICB/AT, RT2 N = 12, RT2.Stat1^−/− N = 10). The mice were sacrificed after 4 weeks of treatment for ex vivo analysis. d, e Representative triple-staining for the senescence marker p16^Ink4a (red) and the proliferation marker Ki67 (blue) and for nuclei (green) (d), e showing quantification of p16^Ink4a+ (left) or Ki67⁺ (right) cancer cells of individual mice, treated as described in a, each data point represents the total of three tumour slides measurements (Ctr, RT2 N = 9, RT2.Stat1^−/− N = 4; AT, RT2 N = 12, RT2.Stat1^−/− N = 5; ICB, RT2 N = 9, RT2.Stat1^−/− N = 3, ICB/AT, RT2 N = 13, RT2.Stat1^−/− N = 7), box plots show the median with 25th and 75th interquartile range (IQR), and whiskers indicate 1.5 × IQR. Significance tested by using Log Rank test (a, left), Fishers exact test (a, right), or two-tailed Mann-Whitney test (c, e). RT2.Stat1^−/− mice have been censored after 3.7 weeks of treatment. Scale bars 200 µm (b) or 2 µm (d). Number of mice is given in parenthesis.