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. 2021 May 6;11:9660. doi: 10.1038/s41598-021-88841-0

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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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Mcu is not required for the development of hair cells. Results obtained in vitro demonstrate that mechanotransduction, Ca²⁺ levels, and mitochondria superoxide activity are not altered in the cochlear hair cells of Mcu^−/− mice. (A) Representative images show FM1-43 uptake in hair cells of Mcu^+/+, Mcu^+/− or Mcu^−/− mice (mid-basal turn). (B) The fluorescence intensity of FM1-43 in hair cells is not significantly different between the groups, indicating the normal functioning of MET channels in Mcu^−/− mice. (C) Representative images showing the MitoSOX indicator levels in hair cells of Mcu^+/+or Mcu^−/−. Asterisks show Mcu^+/+ and Mcu^−/− OHCs, which were excluded from statistical analysis due to no detectable superoxide activity. (D) The fluorescence intensity of MitoSOX in hair cells of Mcu^−/− mice is similar to that of the Mcu^+/+ mice indicating that there is no difference in oxidative activity caused by cellular superoxide buildup. (E,F) The fluorescence intensity of intracellular (Fluo-2 AM) and mitochondrial (Rhod-2 AM) Ca²⁺ indicators in hair cells of Mcu^+/+, Mcu^+/− and Mcu^−/− mice were not significantly different. Data represented herein are from mice of age P5–P9. Two-way ANOVA analysis did not show any statistically significant differences between Mcu^+/+, Mcu^+/− or Mcu^−/− groups. The number of mice for all groups was greater than 5. Scale bars in (A) and (C) are 12 µm.