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. 2020 Sep 2;9(9):2018. doi: 10.3390/cells9092018

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© 2020 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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BFCNs were positive for neuronal markers. BFCN cultures were matured for 4 weeks and immunocytochemistry for general neuronal markers and specific cholinergic neuronal markers was performed. (A–F) Representative confocal pictures of the BFCN cultures at week 4 of BFCN maturation with the nuclear marker Hoechst 33,342 in blue. (A) Cholinergic marker ChAT in green. (B) Neuronal marker β-III-tubulin in red. (C) Merged picture of A and B. (D) Cholinergic marker p75NTR in green. (E) Neuronal marker β-III-tubulin in red. (F) Merged picture of D and E. Scale bars = 50 μm.