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. 2020 Dec 2;11(12):1028. doi: 10.1038/s41419-020-03200-7

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© The Author(s) 2020

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. 2 — a After silencing of SKIL in CALU-3 and NCI-H520 cell lines using shRNA, SKIL expression was significantly inhibited in all the four shRNAs (shSKIL#1–4) compared to control shRNA (shNC), while shSKIL#4 showed the lowest SKIL mRNA expression level. b, c MTT assay showed inhibited viability after SKIL was silenced (shSKIL) in CALU-3 and NCI-H520 compared to control (shNC). d, e Colony formation assay showed fewer colonies formed in CALU-3 and NCI-H520 with silenced SKIL expression (shSKIL) compared to control (shNC). f, g Transwell assay showed inhibited cell migration and invasion in the shSKIL group compared to the control group (shNC). h, i Western blot measurement showed decreased SNAIL1, SLUG, and vimentin expression levels, and increased E-cadherin levels in shSKIL group (shSKIL), compared to control group (shNC) in both CALU-3 and NCI-H520. j, k qPCR results showed that mRNA levels of cancer stem cell markers (CD44, CD133, SOX2, OCT3/4, and NANOG) were decreased after SKIL silencing (shSKIL) compared to control (shNC). l Tumorigenesis assay in nude mice using CALU-3 and NCI-H520 showed slower tumor growth rate and smaller size of collected tumor blocks after animal sacrifice in the shSKIL group (shSKIL) compared to the control group (shNC). *P < 0.05, **P < 0.01. Experiments were performed in triplicate.