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. 2018 Mar 6;37(20):2714–2727. doi: 10.1038/s41388-018-0133-3

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License, which permits any non-commercial 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. If you remix, transform, or build upon this article or a part thereof, you must distribute your contributions under the same license as the original. 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-nc-sa/4.0/.

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Fig. 4 — Treatments with EZH2 inhibitors induce NTRK1 expression. NB-39-nu cells were treated with mock (0.1% (v/v) of DMSO), EPZ6438 (1 μM), or GSK126 (1 μM) for 4 days. Results are representative of at least three independent experiments. a Western blotting for H3K27me3 and EZH2 was performed using EPZ6438- or GSK126-treated cells. b, c NTRK1/GAPDH mRNA levels were assessed by semi-quantitative RT-PCR (b) and qPCR (c). d Western blots for NTRK1 in EZH2 inhibitor-treated NB-39-nu cells. e–g Photos (e) of and percentages (f) of neurite-extending NB-39-nu cells treated with EZH2 inhibitor EPZ6438 and/or NGF. Cells were treated with 1 μM of EPZ6438 for 2 days and cultured for 3 more days after the addition of NGFbeta (Sigma^®,100 ng/ml). Neurite extension was assessed as indicated in Methods and GAP43 expression was studied by semi-quantitative RT-PCR (g)