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. 2022 Aug 17;608(7924):795–802. doi: 10.1038/s41586-022-05082-5

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© The Author(s), under exclusive licence to Springer Nature Limited 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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Extended Data Fig. 1 — a, Schematic of breeding, embryonic stem cell engineering, allelic configuration, and staging of KPC^TRE-shRNA ESC PDAC. Dashed line defines tumour mass detected by ultrasound, K, kidney. b, Representative whole mount bright field and fluorescent gross pathology of PDAC arising in context of adjacent premalignant (Pre-M) tissue in KPC^shRenilla and KPC^shp53 mice. c, Survival curve of KPC^{TRE-shRenilla} (n = 33) and KPC^TRE-shp53 (n = 21) mice. Significance of difference in survival curves assessed by log rank (Mantel-Cox) test. d, Kate and GFP flow cytometry of primary cultures of dissociated shRenilla and shp53 KPC^TRE pancreas following PDAC development at indicated passages. e, PCR detection of recombined versus wild type alleles for Kras and p53 in primary cultures from indicated samples at passage 6. f, Representative flow cytometry plot distinguishing single Kate positive (SP) from double Kate/GFP positive (DP) cells after PDAC development in a KPC^shRenilla mouse. g, PCR detection of recombined versus wild type alleles for Kras, p53, as well as shRNA and RIK transgenes in DP and SP cells sorted from KPC^shRenilla mice following PDAC development and cell lines generated from PDAC arising in KPC^shp53 mice. h, (Top) Digital PCR detecting relative levels of recombined conditional p53 allele, WT p53, and GFP targeted CHC cassette and (bottom) Kras^G12D and WT Kras alleles in DP (n = 16) and SP (n = 19) cells sorted from KPC^{TRE-shRenilla} PDAC. i, Relative copy number of chromosome 11 inferred from sparse whole genome sequencing from PDAC arising in KPC^shRenilla and KPC^shp53 mice (n = 4 each). Normalized segment values are centred around a mean value of 1 with segment values below 1 indictive of deletion events. j, Representative, matched immunofluorescence of GFP and mKate and immunohistochemistry of p53 in sequential sections in a KPC^shRenilla PDAC. h, mean ± S.D. Scale bars b, 1 cm, j 50 μm. e and g were repeated at least twice with similar results, and j was repeated 3 times with similar results. Gel source data for e and g, see Supplementary Fig. 1. Gating strategy of d and f, see Supplementary Fig. 2.

Source data