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. 2023 Apr 17;120(17):e2215253120. doi: 10.1073/pnas.2215253120

Fig. 6.

Fig. 6.

Reprogramming and HC-like cell regeneration in a mouse model with HC loss in vivo. (A) A schematic diagram illustrating the experimental procedure that adult C57BL/6J mice were treated by Kanamycin/Furosemide (Kana/Furo) to kill hair cells, with the subsequent delivery of the cocktail (VLFsiFsiM) or vehicle (sterile water plus 0.1%DMSO) into the middle ear space, followed by the injection of Ad.Atoh1.mCherry into the inner ear by cochleostomy. (B) After Kana/Furo treatment, vehicles/Ad.Atoh1-mCherry-treated adult C57BL/6J cochleae showed scarce hair cells (arrows, ESPN+/mCherry+) in the outer hair cell region (OHCr) of the apex-middle turn. (C) After Kana/Furo treatment, cocktail/Ad.Atoh1.mCherry-treated adult C57BL/6j cochleae showed many hair cells (arrows, ESPN+/mCherry+) in the outer hair cell regions (OHCr) from the apex to the midturn. (D) Quantification and comparison of regenerated HC-like cells showed a significant increase in the HC-like cell number in the OHCr in the cocktail-treated samples compared to the vehicle-treated samples. (E) Quantification and comparison showed a comparable HC number in the IHC region in the cocktail-treated, vehicle-treated, and WT control cochlear samples. (F) Quantification and comparison showed significantly more ESPN+/Atoh1mCherry+ Cells in the cocktail-treated than vehicle-treated control cochlear samples. D: Dox; L: LiCl; F: FSK; siF: siFIR; siM: siMxi1. **P < 0.01, ***P < 0.001, ****P < 0.0001, two-tailed unpaired Student’s t test. Error bar, mean ± SEM; N = 5 to 6 in each group. Source data are provided as a Source Data file. (Scale bars, 10 μm.)