Fig. 1: Characterizing different promoters to drive ATOH1 expression and hair cell regeneration in adult mouse utricle ex vivo.

a Utricles from 4-10-week-old mice were treated with gentamicin (1 mM) followed by AAV8-CMV-ATOH1-H2B-EGFP, AAV8-GFAP-ATOH1-H2B-EGFP or AAV8-RLBP1-ATOH1-H2B-EGFP (4 × 10¹¹ vg/mL), then examined at day 12 and day 20. b–i Utricles treated with CMV-ATOH1, GFAP-ATOH1, RLBP1-ATOH1 or gentamicin control for 12 and 20 days. Insets showing the GFP signals. j, k Mean GFP intensity in hair cells and supporting cells after treatment with CMV, GFAP and RLBP1 promoters. l Quantification shows Pou4f3-positive hair cell counts at day 12 and 20 after treatment with CMV-ATOH1, GFAP-ATOH1, RLBP1-ATOH1 or gentamicin control. Over the time, hair cell number significantly increased with CMV-ATOH1 and GFAP-ATOH1, but not RLBP1-ATOH1 treatment. Data shown as mean ± S.D., compared using two-way ANOVA. p = 0.0063 between CMV-ATOH1 and GFAP-ATOH1 at day 12, and p < 0.0001 for all the others in (j), p = 0.0043 between CMV-ATOH1 and RLBP1-ATOH1 at day 12, p = 0.0001 between GFAP-ATOH1 and RLBP1-ATOH1 at day 20 and p < 0.0001 for all the others in (k). p = 0.0074 between CMV-ATOH1 and RLBP1-ATOH1 at day 12, p = 0.0014 and 0.0191 between day 12 and day 20 with CMV-ATOH1 or GFAP-ATOH1 treatment, respectively. p = 0.006 between RLBP1-ATOH1 and gentamicin control at day 20 and p < 0.0001 for all the others in (l). n = 4, 4, 3 for CMV, GFAP or RLBP1-ATOH1 at 12 days and 5, 5 and 6 at 20 days in (j). n = 4, 4 and 4 for CMV, GFAP or RLBP1-ATOH1 at 12 days and 5, 5 and 6 at 20 days in (k). n = 6, 5, 5 and 5 for CMV, GFAP, RLBP1-ATOH1 or gentamicin control at 12 days and 5, 6, 6 and 4 at 20 days in (l). Scale bars = 100 μm in (b–g). Source data are provided as a Source Data file.