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. 2017 Aug 9;8:219. doi: 10.1038/s41467-017-00254-8

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© The Author(s) 2017

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 — Functional neuropeptidergic hypothalamic neurons (iHTNs) can be derived from hiPSC. a iHTN differentiation protocol. b RT-qPCR of hypothalamic and arcuate nucleus-specific genes showing significantly increased expression of the genes at day 40 of differentiation compared with day 0 (*p < 0.05, **p < 0.01). ND not detectable; two-way ANOVA was employed to determine differences within day 0 and day 40 iHTNs. c Measurement of hypothalamus-specific neuropeptide Y (NPY) measured from cell supernatants using ELISA (p < 0.001 determined using paired t-test). d Measurement of hypothalamus-specific α-melanocyte-stimulating hormone (α-MSH) measured from cell supernatants using ELISA (***p < 0.001 determined using paired t-test). e–n Immunopositivity for hypothalamic progenitors and neuropeptidergic markers. o MEA readings of neurons from day 0 as well as day 40 from the same electrode over time showing increased neuronal firing in day 40 neurons. Images and data shown are representative of average results from the two iPSC lines differentiated n = 5 times in independent experiments. Scale bars represent 50 µm. Error bars denote s.e.m