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. 2021 Dec 17;15:784649. doi: 10.3389/fnins.2021.784649

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Copyright © 2021 Popsuj and Stolfi.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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An Ebf binding site is necessary for VAChT/ChAT reporter activity. (A) Mutating the Ebf binding site in the minimal AMG5-specific VAChT/ChAT cis-regulatory element was sufficient to abolish its activity in AMG5. Mutated reporter n = 50, wild-type reporter n = 44. (B) The AMG5-specific VAChT/ChAT reporter can be ectopically activated in ventral papilla cells at 15.5 h post-fertilization (hpf) at 20°C, by overexpressing Ebf there (using Msx > Ebf). No expression is seen in the negative control electroporated with the neutral construct Msx > lacZ instead. Images are confocal Z stack projections. (C) Yet Ebf overexpression in the ventral papilla does not cause ectopic expression of the mutated VAChT/ChAT reporter, suggesting the Ebf binding site is indispensable for Ebf-mediated activation of this element. For each of the three conditions, n = 50.