Fig. 2

The nampt-a morpholino has off-target effects that lead to the maldevelopment of wild-type and mutant nampt-a embryos. A Phenotypes of nampt-a^+/+ and nampt-a^t10pm/t10pm embryos injected with nampt-a morpholino (nampt-a MO). Scale bars represent 100 µm. B Phenotype distributions of nampt-a^+/+ (+ / +) and nampt-a^t10pm/t10pm (-/-) embryos injected with nampt-a MO. Results are a combination of four experimental trials. + / + , n = 366; + / + with MO, n = 232; -/-, n = 351, and -/- with MO, n = 201. C Time-lapse imaging of embryos with severe phenotypes injected with nampt-a MO at 4, 6, 12 and 12.5 h post-fertilization. Scale bar represents 100 µm. D Wild-type nampt-a^+/+ (+ / +) and mutant nampt-a^t10pm/t10pm (-/-) embryos were injected with nampt-a MO. Embryos were assayed at 1 dpf for nampt-b expression by qRT-PCR, and fold change in expression was calculated using the ∆∆C_t method [26]. Each point represents one biological sample of ten embryos pooled for RNA extraction. Each colored line represents the average from separate experimental trial days, and the black line shows the average from all trials. For statistical analysis in (D, E), the Kruskal–Wallis non-parametric test was applied followed by Dunn’s multiple comparison test. E Each point represents NAD⁺ levels in individuals relative to uninjected nampt-a^+/+ embryos. Each colored line represents the average from separate experimental trial days, and the black line shows the average from all trials