Fig. 11

Effects of loss of pax2a, sp5a, and sp5l on otic patterning. (A) Dorsolateral views of pax2a−/−; sp5a−/−; sp5l−/− triple mutants showing expression of otic markers at 24 hpf (as indicated on the left) following treatment with DMSO (control) or BIO from 12 hpf. Scale bar (Aa), 50 ​μm. (B) Dorsolateral views of anti-Isl1/2 stained SAG neurons at 30 hpf (a, b, e, f, i, j) and brn3c:Gfp ​+ ​hair cells at 48 hpf (c, d, g, h, k, l) under the indicated conditions. Ovals delimit the edges of the otic vesicle. Genotypes of +/+ (wt), sp5a−/−; sp5l−/− double mutants (dKO, double knockouts) and pax2a−/−; sp5a−/−; sp5l−/− triple mutants (tKO, triple knockouts) are indicated. (C, D) Quantification of SAG neurons and hair cells in wild-type embryos, sp5a−/−; sp5l−/− double mutants, and pax2a−/−; sp5a−/−; sp5l−/− triple mutants (as indicated by the key at the top) following treatment with DMSO or BIO from 12 hpf, as indicated. Data show the mean number of anti-Isl1/2 stained SAG neurons (C) and phalloidin stained hair cells in anterior/utricular and posterior/saccular maculae (D). Error bars (standard deviations) and sample sizes are shown for each group, and asterisks indicate significant differences from wild-type embryos for each treatment. Note, it was not possible to identify anterior and posterior patches of hair cells in triple mutants, which produced an unbroken swath of hair cells along the AP axis.