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. 2009 Jan 2;5(1):e1000326. doi: 10.1371/journal.pgen.1000326

Figure 9. The brown mutation is recapitulated in Mc1r-MO knockdown experiments and is rescued by the Surface form of Mc1r in the zebrafish.

Figure 9

Single-celled zebrafish embryos were injected with 0.2 µM of a MO-oligonucleotide directed against the first 25 base pairs of the Danio rerio form of Mc1r (see Methods). Compared with embryos receiving a mock injection of 2× 1 nl of Danieaux's solution, embryos receiving MO injections demonstrate significantly reduced pigmentation. This reduced pigmentation phenotype was highly penetrant, occurring in 95.6% of individuals (n = 206) receiving the injection. Rescue experiments were performed in which individuals receiving Mc1r-MO injection were co-injected with in vitro transcribed RNA from Surface, Pachón, or Yerbaniz constructs. Individuals co-injected with Surface Mc1r RNA were rescued from the reduced pigmentation phenotype (86.1%, n = 108). Individuals co-injected with RNA derived from Pachón or Yerbaniz allele failed to rescue the diminished pigmentation phenotype (20.7% in 58 individuals; 7.4% in 54 individuals, respectively). Scale bar: 500 µm.