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. 2022 Jan 20;23(3):1123. doi: 10.3390/ijms23031123

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© 2022 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 (https://creativecommons.org/licenses/by/4.0/).

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EN1 increases the activity of proliferation related Hedgehog signaling pathways. (A) Overrepresented Gene Ontology (GO) terms from RNA−seq analysis of upregulated gene sets and downregulated gene sets in EN1 knockdown group compared to control cells. (B) Immunoblotting assays showing that EN1 knockdown significantly reduces the Gli1 level in glioma cells. (C) The EN1 and Gli1 transcript levels in U251 cells transduced with scramble control, shEN1, shcon + Gli1, or shEN1 + Gli1. (D) Overexpression of Gli1 rescues the restrained colony-forming abilities caused by the knockout of EN1 (□ for U251-sgLacZ, ● for U251-sgEN1, ■ for U251+Gli1-sgEN1). Data are presented as the mean from three independent experiments. * p < 0.05, ^### p < 0.001 according to Student’s t-test.