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. 2017 Nov 29;8:1848. doi: 10.1038/s41467-017-01845-1

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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 Commonslicense, unless indicated otherwise in a credit line to the material. If material is not included in the article’sCreative 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 — Multipotent progenitor and NSC-like profile of LAM and angiomyolipoma. a Hematoxylin and eosin (H&E) of the lung sections from sporadic LAM; GFAP or NSE expression in LAM. Arrows indicate LAM and arrowheads normal bronchial mucosa (upper) or normal pleura (lower images). b Sporadic angiomyolipomas (upper left and upper middle) and adjacent normal kidney (upper right) or TSC-associated (lower images) angiomyolipomas, stained with H&E or for NSE, GFAP, NS-tubulin, or nestin (arrows indicate angiomyolipoma and arrowheads kidney). c (i) GFAP, (ii) NSE and nestin expression in (i) TSC-associated and sporadic (ii) angiomyolipomas (T) from patients (P) vs. normal kidney (NK) by western immunoblotting. d (i) Nestin and peripherin expression in LAM (first left) and co-expression analysis by digital pathology; red indicates peripherin (second), brown nestin (third image), and yellow indicates co-expression of both markers (markup, right fifth panel: details within the area marked by the corresponding insert in fourth panel, black arrows). (ii) Quantification for data shown in d (i) by Aperio Digital Pathology; the percentage of area with co-expression of peripherin and nestin in LAM from 9 patients. Error bars are defined as means + s.e.m. e (i) FACS of cells grown in DMEM medium. Percentages of 621–101 and CRL4004 (angiomyolipomas-derived) cells expressing nestin alone or co-expressing: melan-A, GFAP, SMA, and NS-tubulin; (ii) Nestin, GFAP, and NS-tubulin expression in 621–101 (grown in DMEM or IIA complete medium), CRL4004 angiomyolipoma cells, angiomyolipoma tumor, and corresponding normal kidney by western immunoblotting. Error bars are defined as means + s.e.m. f q(RT)-PCR of SOX10, SOX9, ID3, NESTIN, c-Kit, DCT, and S100A1 mRNA levels relative to 18S in 621–101, CRL4004, angiomyolipoma tumor, and corresponding normal kidney. Data represent means (±s.e.m.). Data from nine LAM and eleven angiomyolipomas (a–d). Scale bar: 50 µm. *P ≤ 0.05, t-test. Data are representative of three to six (e (i)) independent experiments