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. 2017 Sep 26;7:12317. doi: 10.1038/s41598-017-12017-y

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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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IL-6 plays an important role in the formation of lung cancer organoids. (A) Phase contrast and fluorescence images of the organoids consisting of EGFP-labeled parental A549 cells, HUVECs and MSCs. (B) The mean fluorescence intensity of GFP in the EGFP-labeled parental A549 cell-derived organoids. The addition of IL-6 significantly strengthened the mean fluorescence intensity (n = 3, **P < 0.01; two-tailed paired t-test). (C) HE and αSMA staining showed that the addition of IL-6 (10 ng/ml) increased the organoid forming ability of A549 + HUVEC + MSC by collaborating with the αSMA-positive cells. (D) The proportion of αSMA-positive cells in the parental A549 derived-organoids was increased by the addition of IL-6 (10 ng/ml) (n = 3, **P < 0.01; two-tailed paired t-test). (E) Phase contrast and fluorescence images of the organoids consisting of EGFP-labeled OSK-A549-colony cells, HUVECs and MSCs. (GFP shows cancer cells). (F) The mean fluorescence intensity of GFP in the EGFP-labeled OSK-A549-Colony cell-derived organoids. The addition of IL-6 antibodies did not attenuate the mean fluorescence intensity (n = 3, two-tailed paired t-test). (G) αSMA and HE staining showed that IL-6 signal blockade attenuated the organoid construction ability of OSK-A549-Colony + HUVEC + MSC, and that the proportion of αSMA-positive cells in the organoids was decreased. (H) The proportion of αSMA-positive cells in the organoids was decreased by the addition of IL-6 antibodies (n = 3, **P < 0.01; two-tailed paired t-test). (I) Immunocytochemistry for αSMA. Seven days treatment of IL-6 (10 ng/ml) could induce αSMA-positive cells from MSCs culture.