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. 2017 Nov 13;8:1466. doi: 10.1038/s41467-017-01514-3

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© The Author(s) 2017

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. 1 — An ex vivo genetically engineered model of renal AML. a Proliferation assay, western blot and soft-agar assay of Cdkn2a^∆/∆ primary kidney cells following infection with lentiviruses expressing an empty shRNA (shRNA-Ctrl) or shRNAs against Tsc1 (shRNA-Tsc1) or Tsc2 (shRNA-Tsc2). b Proliferation assay, western blot and soft-agar assay of primary kidney cells infected with lentiviruses expressing shRNAs against Cdkn2a (shRNA-Cdkn2a) or Tsc2 and Cdkn2a (shRNA-Tsc2 + Cdkn2a). All graphs depict mean ± s.d. Student’s t test, n = 3. ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001. c Phase contrast images of spheres formed in suspension culture conditions from shRNA-Tsc2/Cdkn2a^∆/∆ and shRNA-Tsc2 + Cdkn2a primary kidney cells. Scale bars: 50 μm. d Representative allografttumours derived from subcutaneous injections of respective sphere cells into SCID-beige mice.Scale bars: 1 cm. e Representative histology of allograft renal AML-like tumours. Characteristic histological features of AML are indicated by letters, where A shows adipocyte, S shows spindle cells and V shows an aberrant blood vessel. Epithelial structures are indicated by Ep. Scale bars: 50 μm. f Positive immunofluorescence or immunohistochemical staining in engineered renal AML-like allograft tumours for PMEL(HMB45), HMB45/MART-1, CATHEPSIN K, SMA, VIMENTIN, PDGFRβ, CD31 and S100. Scale bars: 50 μm