Zebrafish assessment
Live embryo phenotypes were analyzed using a Leica MZ12.5 at 24 and 48hpf. The pattern of rescue was determined by the somite shape and eye color in the rescue experiment. We determined the phenotypic difference in rescue experiments based on specific abnormal features of smoK/O model: U -type tail and less than half area of retina pigmentation in the eye region. Embryo phenotypes with partial rescue of eye shown over half area of retina pigmentation. Embryo phenotypes with partial rescue of somite shown not curled U -type tail but rather an unstraight tail. Live embryo images of lateral or dorso-ventral views, images of gene expression after in situ hybridization and images for eye area or somite analysis were acquired on a ZEISS AxioCam HRC Camera and AxioVision SE64 software after embedded embryos in 3% Methylcellulose (Sigma): Live embryo and eye area or somite analysis; in Glycerol (Invitrogen): in situ hybridization. Somite angle measurements were taken at the midpoint between the proximal hindgut and the anus at 24hpf embryos using ImageJ software. For eye area measurements, lateral images at 24hpf embryos were quantified using ImageJ software. For eye and somite analysis, total 45 embryos/group: 15 embryos/group in each triplicate injection, were analyzed.