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. 2020 Jun 4;11:2817. doi: 10.1038/s41467-020-16309-2

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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. 2 — a β-galactosidase staining of pancreatic sections from AGO2^+/+;KRAS^G12D;p48Cre and AGO2^fl/fl;KRAS^G12D;p48Cre mice at 70- and 500-day time points. Scale bar, 100 µm. b Scatter plot showing β-galactosidase staining in low grade PanINs. Data are from 47 PanINs from AGO2^+/+;KRAS^G12D;p48Cre and 98 PanINs from AGO2^fl/fl;KRAS^G12D;p48Cre from four individual mice at the 70-day time point and 30 PanINs from three individual mice at 500-day time points. Intensity of staining and percent cells within 30 low grade PanINs were used to determine the senescence score = intensity × percent positive cells. p values were determined using a two sided t-test. Data are presented as mean values +/− SEM. c Immunoblot analysis of RAS-driven MAPK (indicated by pERK) and PI3K (indicated by pAKT) signaling from individual pancreata obtained from mice of the indicated genotypes, aged to 400 days. Numbers on the left indicate protein molecular weights in kDa. d Representative images of H&E staining (left) and dual staining for β-galactosidase and phospho-ERK (right) in human pancreatic tissue with PanINs (representative staining of at least 10 PanINs from two patients). Scale bar, 40 µm. e Immune profile of lesions from the indicated genotypes. Ten consecutive fields (20x magnification) from four individual mice were assessed for the indicated IHC markers that distinguish immune cell populations. Significant p values are indicated and were determined using two tailed t-test. f Representative images of NK1.1-positive NK cells surrounding PanIN and PDAC lesions within the indicated genotypes. Scale bar, 50 µm. g Plot showing NK cell number in PanIN/PDAC lesions within the indicated genotypes. Pancreatic tissues from six mice were analyzed for NK1.1 IHC-positive NK cells. Two tailed t-test was used to determine p values. h Scatter plot showing peripheral and PanIN infiltrating NK cell count from PanINs in the indicated genotypes. Counts were obtained from 10 consecutive fields from six mice at ×20 magnification, and the indicated p values were determined using two tailed t-test. In relevant panels, data are presented as mean values +/− SEM.