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. 2017 Dec 21;10:426. doi: 10.3389/fnmol.2017.00426

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Copyright © 2017 Chen, Lu, Guo, Ni, Zhang, Zhao, Wu, Sun, Zhang, Tang, Li, Chai and Li.

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) or licensor 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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Forced activation of Hedgehog signaling did not trigger cell proliferation or new HC formation in the cochlear epithelium without neomycin treatment. (A) The diagram for the assay. Cochlear epithelium samples were dissected from P2 Smo-OE and control mice and then cultured in DMEM/F12 media with N2 and B27. 4-OH tamoxifen (Tm) and EdU were added throughout the culture period. At the end of 7 days of culture, the tissues were fixed and immunostained for Myo7a, Sox2 and EdU. (B,C) Representative immunofluorescence images of the supporting cell layer in the control and Smo-OE cochlear epithelia without neomycin treatment. No difference in the number of EdU+Sox2+ cells was seen between the Smo-OE and control groups. (D,E) Representative immunofluorescence images of the HC layer in the control and Smo-OE cochlear epithelia without neomycin treatment. No EdU+Myo7a+ HCs were observed in either group. Scale bars (B–E) are 20 μm.