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. 2017 Apr 3;36(32):4641–4652. doi: 10.1038/onc.2017.91

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Copyright © 2017 The Author(s)

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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GLI1 inhibition diminishes proliferation and stemness in LAC cells. (a–d) LAC cells viability is diminished by GLI1 knockdown in (a) commercial cell lines (siRNA-mediated) and (b) patient-derived CSCs (shRNA-mediated); similar reductions are seen in (c) cell lines and (d) CSCs treated for 48 h with the GLI1 antagonist GANT61 or DMSO (−). Bar graphs show viability (MTS assay); immunoblots show endogenous GLI1 protein levels. LC: GAPDH. Numbers below blots indicate densitometrically quantified protein expression. (e) Effects of GANT61 on oncosphere formation in LAC CSC lines before and after transduction with shGLI1. Bar graphs: frequencies of oncosphere-forming cells (OFC, % of seeded cells that formed oncospheres) under indicated experimental conditions are normalized to the frequency observed in shSCRAMBLE-infected cells treated with DMSO alone (controls, CTRL, expressed as 100%). Photomicrographs: Representative images of oncospheres observed in DMSO-treated CSCs infected with shGLI1 or shSCRAMBLE (CTRL). Scale bar: 50 μm. (f) Percentage of CSCs exhibiting high ALDH activity (ALDH⁺ cells) after GANT61 or CTRL treatment (AldeFluor assay). (g) Expression (mRNA and protein) of stem cell markers OCT4 and ABCG2 in GANT61- and CTRL-treated (dashed line) CSCs. LC: GAPDH. *P< 0.05. ALDH, aldehyde dehydrogenase.