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. 2021 May 28;11:11295. doi: 10.1038/s41598-021-90798-z

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Generation of iPSC and NPC lines from individuals with MAND. (a) Bright-field images of undifferentiated iPSC lines from MAND1-3 (top row panel, l to r). Lower panel images are three NPC lines derived from MAND1-3 iPSC lines, respectively (l to r). After two weeks of neuronal induction, images were captured. Bars, 0.2 mm. (b) Immunostaining of MAND1 iPSC (1^st row) and MAND1 NPC (2^nd row) for neural progenitor cell marker PAX6 (1^st column) and pluripotent cell marker NANOG (2^nd column and 3^rd for merged). Immunostaining of MAND1 iPSC and MAND1 NPC for NES (3^rd and 4^th rows, respectively). (c, d) qRT-PCR analysis confirmed ~ 50% reduction of MBD5 mRNA expression in MAND iPSC (c) and NPC lines (d) when compared with control iPSC and NPC lines, respectively. p < 0.0001. Quantification of the test gene (MBD5) was calculated relative to that of the housekeeping gene, GAPDH. All expression values were calculated relative to control levels set at 1.0. Error bars represent the SEM. ****p < 0.0001, n = 3 independent biological replicates performed in triplicate, GraphPad Prism 8.4.2 (graphpad.com).