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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 promoted in situ supporting cell proliferation and HC regeneration after 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. The explanted cochleae were treated with 0.5 mM neomycin sulfate (Neo) for 24 h. 4-OH tamoxifen (Tm) and EdU were added to the culture media throughout the entire culture period to induce the Cre activity and to label proliferating cells, respectively. (B,C) Representative immunofluorescence images of the supporting cell layer in the control and Smo-OE cochlear epithelia after neomycin treatment. (D,E) Representative immunofluorescence images of the HC layer in the control and Smo-OE cochlear epithelia after neomycin treatment. (F) The number of EdU+Sox2+ cells significantly increased in the Smo-OE group compared with the control group. (G) The number of EdU+Myo7a+ cells significantly increased in the Smo-OE group compared with the control group. (H) The ratio of EdU+Myo7a+ cells to EdU+Sox2+ cells in the Smo-OE and control groups. (I) The total number of Sox2+ supporting cells (SCs). Data are presented as means ± SE. N = 5, *p < 0.05 vs. the control group with neomycin treatment. Scale bars (B–E) are 20 μm.