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. 2017 Jul 6;7:4770. doi: 10.1038/s41598-017-05047-z

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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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RA treatment induces TKI resistant NSCLC cell differentiation. (a) qRT-PCR analysis of GATA6 and NKX2.1 in H-1975 NSCLC cell line treated with 1 and 10 µM RA for 24 and 48 hrs. The comparison has been conducted by using the ΔΔCT method and normalized to GAPDH transcript. Dotted line represents the normalized expression levels of each transcript analyzed in untreated cells. Data are represented as mean ± SD. Statistical analysis has been obtained by paired t test (n = 3). (b) Left panel, Representative Western blotting analysis of GATA6 and GAPDH in H-1975 cells treated with 1 and 10 µM RA for 24 and 48 hrs. Right panel, densitometric analysis of GATA6 levels, normalized versus GAPDH, used as loading control (n = 4) (*=0.02). (c) qRT-PCR analysis of GATA6 target genes in H-1975 NSCLC cells treated with 1 and 10 µM RA for 48 and 72 hrs. The comparison has been conducted by using the ΔΔCT method and normalized to GAPDH transcript. Dotted line represents the normalized expression levels of each transcript analyzed in untreated cells. Data are represented as mean ± SD. Statistical analysis has been obtained by unpaired t test (n = 3).