Figure EV5. Wrb disruption affects IHC morphology.

• A, A′
  Representative low‐magnification (10×) confocal projections of apical turn whole‐mount organs of Corti from Wrb ^+/+:Cre ^A P17 (A, same specimen as shown in Appendix Fig S4) and Wrb ^fl/fl:Cre ^A P21 (A′) animals immunolabeled for GFP (green, reporting Cre recombination), otoferlin (magenta, labeling IHCs), and DAPI (blue, labeling all cell nuclei). Scale bar: 100 μm. Note that Wrb deficiency does not lead to a dramatic loss of IHCs, but a change in IHC morphology. Cell shape and nuclear position were analyzed in images taken with higher magnification (63×, see Fig 4).
• B, C
  Wrb‐deficient IHCs (P18–21) show significantly decreased sagittal, but increased coronal extent (Wilcoxon rank test, ***P < 0.001), resulting in an increased ratio of these two measurements (Wilcoxon rank test, ***P < 10⁻¹⁵, n = 46 and 67 Wrb‐deficient and control IHCs from four and six P18–21 Wrb ^+/+:Cre ^A and Wrb ^fl/fl:Cre ^A animals, respectively). Custom‐written MATLAB routines were used for the analysis. Data are represented as means ± SEM.
• D
  Nuclear position is altered in Wrb ^fl/fl:Cre ^A animals, where nuclei are located closer to the cell base as compared to controls. The relative distance from the apical plane of a cell is given. Data are represented as means ± SEM.
• E, F
  Schematic drawings illustrating the analysis performed in (B–D).