Fig. 3: Trafficking of MYO15A and elongation complex in jordan hair cells.
A, B Immunofluorescence (IF) confocal images showing anti-MYO15A (green, PB48) in control Myo15a+/jd, Myo15ash2/sh2, and Myo15ajd/jd IHCs fixed at P14 (A), and Myo15a+/jd and Myo15ajd/jd IHCs at P7 (B). Phalloidin was used to label F-actin (magenta). Strong extra-cellular PB48 labelling was observed independent of genotype at P7 and thought to be artefactual (asterisk). C Quantification of MYO15A antibody (PB48) at the tips of row 1 stereocilia in P7 IHCs. N = 29 hair bundles (+/jd) and n = 29 hair bundles (jd/jd). Three independent mice were quantified per condition. D–G IF confocal images of elongation complex proteins (green) WHRN (D), EPS8 (E), GPSM2 (F) and GNAI3 (G) in control Myo15a+/jd and Myo15ajd/jd IHCs fixed at P7, overlaid with phalloidin labelled F-actin (magenta). H–K P7 quantification of IF labelling at row 1 stereocilia tips for WHRN (+/jd: n = 20, jd/jd: n = 25) hair bundles (H), EPS8 (+/jd: n = 20, jd/jd: n = 20) hair bundles (I), GPSM2 (+/jd: n = 12, jd/jd: n = 11) hair bundles (J) and GNAI3 (+/jd: n = 12, jd/jd: n = 12) hair bundles (K). Two (GPSM2 + GNAI3) and three (WHRN + EPS8) independent mice quantified per condition. Images are representative of data from at least two independent animals per genotype / antibody. Images comparing antibody labelling between +/jd and jd/jd genotypes are mapped equally. Data are mean ± SD, ****, P < 0.0001, ***, P < 0.001, **, P < 0.01, computed using a Mann-Whitney U-test.
