Fig. 5. Morpholino knock-down of Shroom2 results in severe RPE hypopigmentation. (A) RT-PCR confirmation of morpholino efficacy. Normally spliced Shroom2 mRNA was absent from 80 ng Shroom2-SPL-MO injected embryos. (B) Shroom2-SPL-MO or Shroom2-5MM-MO (80 ng) was injected into two animal blastomeres at the four-cell stage and embryos were grown to stage 42. (C,D) Shroom2-SPL-MO injected embryos show severe reduced RPE pigmentation (E,e') when compared with stage-matched uninjected or Shroom-5MM-MO embryos (C-d'). (F) Measurement of pixel density in the eye indicates no significant difference in pigmentation between uninjected and Shroom2-5MM-MO-injected embryos at stage 39, but a drastic difference between pixel density between controls and Shroom2-SPL-MO eyes.