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. 2017 Feb 24;18(3):486. doi: 10.3390/ijms18030486

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© 2017 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Inhibiting SMO pharmacologically and genetically leads to dysregulation of NANOG expression in vitro model. (A) A co-culture model of an AML cell line (THP-1) with stroma which overexpresses human sonic hedgehog (SHH) ligands; (B) NANOG transcripts substantially decreased during PF-913 treatment in THP-1 cells co-cultured with MS-5 cells over-expressing human SHH ligands. In a pre-clinical setting, although the level of GLI-target gene transcript did not decrease much, that of NANOG decreased during the time course; (C) Western blotting showed the NANOG protein level decreased during PF-913 treatment; (D) Western blot analysis showed downregulated NANOG protein after transfection with SMO siRNAs in comparison to cells transfected with scrambled control. Although knockdown effect of SMO by si-SMO#1 was weak compared with #2 or #3, NANOG expression was evident as well.