Fig. 5: POU4F3 nuclear localization restoration and hair cell survival improvement in Pou4f3^WT/Q113* mice post-injection.

A Representative immunofluorescence images of POU4F3 localization in OHCs and IHCs across cochlear regions (apex, middle, and base) in WT, Pou4f3^WT/Q113*, and AAV-treated Pou4f3^WT/Q113* mice, n = 3 independent experiments with similar results. Dashed circles represented the edges of hair cells. Myo7a (green): hair cell marker. Scale bar: 20 μm (top and middle rows), 5 μm (bottom row). B The proportion of POU4F3 protein signal overlapping the nuclear signal in the apical, middle, and basal turns of WT (gray), Pou4f3^WT/Q113* (blue), and AAV-treated Pou4f3^WT/Q113* mice (red) at P90. In OHCs (n = 60), WT vs Pou4f3^WT/Q113*, p < 0.000001 (apex, middle, and base), Pou4f3^WT/Q113* vs Pou4f3^WT/Q113* + AAV, p = 0.000455 (apex), p = 0.000004 (middle), p = 0.014995 (base); WT vs Pou4f3^WT/Q113* + AAV, p = 0.023242 (apex), p = 0.125571 (middle), p = 0.003470 (base). In IHCs (n = 30), WT vs Pou4f3^WT/Q113*, p < 0.000001 (apex, middle, and base), Pou4f3^WT/Q113* vs Pou4f3^WT/Q113* + AAV, p < 0.000001 (apex, middle, and base); WT vs Pou4f3^WT/Q113* + AAV, p = 0.342315 (apex), p = 0.012933 (middle), p = 0.000280 (base). C The proportion of edited/total OHCs or IHCs in 100 μm cochlear segments of the apical, middle, and basal turns for WT (gray), Pou4f3^WT/Q113* (blue), and AAV-treated Pou4f3^WT/Q113* mice (red) at P90 on enumeration of OHCs/IHCs with restored nuclear POU4F3 signals. In OHCs (n = 5), WT vs Pou4f3^WT/Q113*, p < 0.000001 (apex), p = 0.000001 (middle), p < 0.000001 (base); Pou4f3^WT/Q113* vs Pou4f3^WT/Q113* + AAV, p = 0.003261 (apex), p = 0.002853 (middle), p = 0.003530 (base); WT vs Pou4f3^WT/Q113* + AAV, p = 0.004828 (apex), p = 0.041546 (middle), p = 0.005185 (base). In IHCs (n = 5), WT vs Pou4f3^WT/Q113*, p < 0.000001 (apex and middle); Pou4f3^WT/Q113* vs Pou4f3^WT/Q113* + AAV, p < 0.000001 (apex), p = 0.000048 (middle), p = 0.000018 (base); WT vs Pou4f3^WT/Q113* + AAV, p = 0.013850 (apex), p = 0.092971 (middle), p < 0.000001 (base). D Representative immunofluorescence images of the cochleae in the apical, middle, and basal turns from untreated or AAV-treated Pou4f3^WT/Q113* ears, n = 4 independent experiments with similar results. Scale bar, 20 μm. E Quantitative comparison of surviving OHCs and IHCs in the apical, middle, and basal turns from uninjected or AAV-injected Pou4f3^WT/Q113* ears, OHC (n = 4), Pou4f3^WT/Q113* vs Pou4f3^WT/Q113* + AAV: p = 0.113532 (apex), p = 0.000518 (middle), p = 0.000017 (base); IHC (n = 4), Pou4f3^WT/Q113* vs Pou4f3^WT/Q113* + AAV, p = 0.355918 (apex), p = 0.704853 (middle), p = 0.207031 (base). Data in (B, C, E) are represented as mean ± SEM. Statistical significance was determined by two-tailed Student’s t-tests (^*p < 0.05, ^**p < 0.01, ^***p < 0.001, ns no significant difference). Source data are provided as a Source data file.