wnt11 (wnt11/1) is expressed in the lateral line, interacts with gpc4 and does not act redundantly with wnt11/2 in setting up hair cell orientation. a. Priml-derived neuromasts are labeled with black arrowheads. White arrowheads indicate Primll-derived neuromasts that are outlined by using a dotted line. Priml-derived neuromasts show wnt11 (wnt11f1) mRNA expression on the anterior side but no signal is present in primll-derived neuromasts {see also Fig. 1d). b-n. Analysis of hair cell orientation with respect to the nearest neighbor. b. Diagram showing how the angle between two neighboring hair cells was calculated based on their relative positions to each other. For more details, see Methods. c-n. Histograms of angular data and fitted distribution (red) for each of the conditions analyzed. c-h. Binned angle distribution in priml-derived neuromasts. The von Mises distribution is shown in red to display the distribution of the data. c. Distribution in priml-derived neuromasts of wild type fish. Note the tight degree of alignment between neighbors (Uniform distribution p-value=7.7S x 10^-107). d-f. Distribution in priml-derived neuromasts of Wnt pathway mutants. They all show a high degree of alignment (Uniform distribution p-value in c= 1.21 x 10^-277, d= 1.97 x 10^-184 and e=S.46 x 10^-44). g. Angle distribution in priml-derived neuromasts of vang/2 mutants. Neighbor hair cells do not show angle coordination (Uniform distribution p-value= 0.212). h. Angle distribution in priml-derived neuromasts of MZscrib mutants. Hair cells show a high degree of coordination between neighbors (Uniform distribution p-value= 7.11 x 10^-86), but the distribution is more dispersed than the one shown by wild type fish. i-n. Binned angle distribution in primll- derived neuromasts. i. Angle distribution in primll-derived neuromasts of wild type fish. Note the high degree of alignment between neighbor hair cells (Uniform distribution p-value= 2.08 x 10^-106). j-l. Angle distribution in primll-derived neuromasts of Wnt pathway mutants. ln all three conditions, hair cells show significant degree of coordination between neighbors (Uniform distribution p-value in i= 3.09 x 10^-18, j= 1.97 x 10^-07, k= 7.03 x 10^-14). m. Angle distribution in primll-derived neuromasts of vang/2 mutants. Neighbor hair cells do not show coordination in their alignment (Uniform distribution p-value= 0.0S9). n. Angle distribution in primll- derived neuromasts of MZscrib mutants. Hair cells show a high degree of coordination between neighbors (Uniform distribution p-value= 1.09 x 10^-33), but the distribution is more dispersed than the one shown by wild type fish. o. Hair cell orientation is disrupted only in primll-derived neuromasts of 5dpf ( MZwnt11wnt11f1) homozygous siblings and MZwnt11 (wnt11f1);gpc4 double homozygous fish. Double MZwnt11 (wnt11f1);gpc4 mutants do not show a hair cell phenotype in priml neuromasts. p. Hair cell orientation in 5dpf wnt11f2 homozygous, wnt11 (wnt11f1) homozygous and double wnt11f2;wnt11 (wnt11f1) homozygous fish. Only a mutation in wnt11 (wnt11f1) disrupts hair cell orientation in primll-derived neuromasts. Thus, wnt11 (wnt11f1) and wnt11f2 do not act redundantly during the establishment of hair cell orientation. The cell axis defined by the position of the kinocilium is labeled in yellow. Scale bar in a equals 50µm, scale bar in o and p equals 5µm.
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