Fig. 1

EpCAM regulates posterior lateral line neuromast development and regeneration. (A-B) Whole mount images of 5pdf wild-type and mzepcam mutant zebrafish larvae showing expression of epcam mRNA by in situ hybridization. Scale bar=100?m. Dashed squares show area of individual neuromasts in (A?-B?). Scale bar=20?m. (C-F) RNA in situ hybridization of epcam at 5dpf pre-NEO exposure, 3-hours post-NEO exposure, 1-day post-NEO exposure at 6dpf, and 3-days post-NEO exposure at 8dpf. Scale bar=20?m. (G-H) Live confocal projections of 5pdf wild-type and mzepcam mutant larvae showing hair cells labeled with DASPEI in the primary posterior lateral line neuromasts (L1-L5) and terminal cluster (tc) neuromasts. Scale bar=100?m. (I) Quantification of DASPEI-labeled posterior lateral line neuromasts in 5pdf wild-type and mzepcam mutant zebrafish. n=8 larvae per condition. Data presented as ± S.D. Mann-Whitney test, significance at p<0.05. (J-K??) Confocal projections of neuromasts in 5pdf wild-type and mzepcam mutant larvae with hair cells labeled with a-Oto antibody and nuclei labeled with DAPI. Scale bar=20?m. (L-M) Quantification of ?-Oto-labeled hair cells and DAPI total cells in neuromasts of 5pdf wild-type and mzepcam mutant larvae. n=29 neuromasts in 9 wild-type and n=27 neuromasts in 10 mzepcam mutant larvae. Data presented as ± S.D. Mann-Whitney test. Data presented as ± S.D. Mann-Whitney test, significance at p<0.05. (O-P??) Confocal projections of neuromasts in 8pdf wild-type and mzepcam mutant larvae with hair cells labeled with ?-Oto antibody and nuclei labeled with DAPI. Scale bar=20?m. (Q-R) Quantification of ?-Oto-labeled hair cells and DAPI total cells in neuromasts of 8pdf wild-type and mzepcam mutant larvae. n=32 neuromasts in 10 wild-type and n=27 neuromasts in 10 mzepcam mutant larvae. Data presented as ± S.D. Mann-Whitney test. Data presented as ± S.D. Mann-Whitney test, significance at p<0.05. (S-T??) Confocal projections of neuromasts in 8pdf following hair cell ablation with NEO at 5dpf in wild-type and mzepcam mutant larvae with hair cells labeled with ?-Oto antibody and nuclei labeled with DAPI. Scale bar=20?m. (U-V) Quantification of regenerated ?-Oto-labeled hair cells and DAPI total cells in neuromasts of 8pdf wild-type and mzepcam mutant larvae. n=33 neuromasts in 11 wild-type and n=32 neuromasts in 11 mzepcam mutant larvae. Data presented as ± S.D. Mann-Whitney test. Data presented as ± S.D. Mann-Whitney test, significance at p<0.05. Abbreviations: days post fertilization (dpf), neomycin (NEO), standard deviation (S.D.), ?-Otoferlin antibody (?-Oto).