Fig. 7: Reduced responsiveness of gemin5^hg81 mutants to the knockdown of ErbB pathway genes.

a Quantification of lateral line neuromasts in the mock and AG1478-treated erbb3b^hg115 mutant embryos at 5 dpf. b Quantification of lateral line neuromasts in the mock and AG1478-treated nrg1^hg114 mutant embryos at 5 dpf. Approximately 40 embryos generated from a pair of heterozygous parents carrying either erbb3b^hg115 or nrg1^hg114 mutation were treated with 0 or 2 µM AG1478 from 1 to 5 dpf and then stained with Yopro-1 to count lateral line neuromasts. c Quantification of lateral line neuromasts in the gemin5^hg81/erbb3b^hg115 mutant embryos at 5 dpf. The data are generated from analyzing 177 embryos generated from pairwise incrosses of double heterozygous parents and five embryos are double mutants. d Quantification of lateral line neuromasts in the gemin5^hg81/nrg1^hg114 mutant embryos at 5 dpf. The data are generated from analyzing 156 embryos generated from pairwise incrosses of double heterozygous parents and 13 embryos are double mutants. e erbb2 mutation rate in erbb2 CRISPR guide RNA injected gemin5 mutant embryos at 5 dpf. Mutation rate was measured by CRISPR-STAT fluorescent PCR-based fragment analysis⁶¹. f Quantification of lateral line neuromasts in the mock and erbb2 CRISPR guide RNA injected gemin5 mutant embryos at 5 dpf. Error bars in the graphs indicate mean ± s.e.m. ns, P > 0.05; *P < 0.05; **P < 0.01; ****P < 0.0001. The analysis was done by injecting the embryos generated from a pair of gemin5^hg81 heterozygous parents with either Cas9 protein alone (mock injection) or Cas9 protein together with erbb2 guide RNAs (erbb2 gRNA injection), followed by analyzing hair cell development in ~40 injected embryos for each condition, and lastly determining gemin5^hg81 genotype and erbb2 mutation rate for each of the analyzed embryo. No erbb2 mutation was detected in the mock injection group.