FIGURE 4

Papp-aa and Igfbp5a are indispensible for Stc1a action in NaR cells. (A) Embryos of the indicated genotypes were raised in E3 embryo medium until 5 dpf. The mRNA levels of papp-aa were measured and normalized. n = 15–17 larvae/group. (B)Tg(igfbp5a: GFP) embryos were injected with stc1a targeting gRNAs and Cas9 mRNA at the 1-cell stage. Embryos were raised in E3 embryo medium. The injected embryos were treated with BMS-754807 (BMS, 0.3 μM), ZnCl₂ (8 μM), or Bastimastat (200 μM) from 3 to 5 dpf. NaR cells were quantified and shown. n = 20–39 larvae/group. (C,D) Progeny of papp-aa^+/–; Tg (igfbp5a: GFP) intercrosses (C) or igfbp5a^{– /–}; Tg (igfbp5a: GFP) intercrosses (D) were injected with stc1a targeting gRNAs and Cas9 mRNA. The injected embryos were raised and NaR cells were quantified at 5 dpf. Each larva was genotyped afterward. n = 5–19 larvae/group (C) and n = 39–43 larvae/group (D). (E,F) Larvae of the indicated genotypes were raised in E3 embryo medium and NaR cells were quantified at 5 dpf. n = 9∼78 larvae/group (E) and n = 4∼34 larvae/group (F).