SEM images from the 1/4 apical turn of the cochlea of P10, P12, and P14 Strc+/−/Tecta−/− and Strc−/−/Tecta−/− mice were analyzed using ImageJ software [National Institutes of Health (NIH), Bethesda, MD, USA] to measure the stereocilia hair bundle effective area at the top of the bundle (Ab). We used the selection brush tool to generate a yellow contour (fig. S2) around stereociliary bundles and determined the area under the contour. Last, we defined three separate groups: P9 and P10 mice with an effective area equal to measurements at P10, intermediate P11 and P12 mice with an effective area equal to measurements at P12, and P13 to P15 mice with an effective area equal to measurements at P14. The reasoning behind the generation of groups was the development of horizontal top connectors. Around P11 is when stereocilin appears in the cochlea apical coil turn and progressively matures (14). Therefore, we believe that it is adequate to assume that, from P13 to P15, the effective areas are equal because of the presence of horizontal top connectors.

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