Morpholino-mediated depletion of Lzap results in cleavage stage and epiboly defects. A: Schematic representation of lzap RNAs. lzap:3HA contains the 5′ untranslated region (UTR) sequence targeted by the lzap morpholino (MO) and was used to confirm MO effectiveness. lzap:CDs contains the entire lzap coding sequence but lacks the MO target sequence and was used for rescue. B: Verification of on-target and dose-dependent effects of lzap MO on exogenous Lzap expression. Embryos were injected with lzap:3HA singly or with increasing amounts of lzap MO visualized by immunofluorescence 4 hours postfertilization (hpf). No immunofluorescence was observed in control embryos lacking the primary antibody (data not shown). C: Schematic showing incidence of developmental defects and nonviable embryos at indicated time points following injection of lzap MO, control MO or in embryos without injection. D–E′′: lzap specific MO or control was injected and viability and morphology determined by microscopic visualization at indicated times. Results were compared with uninjected embryos. D–H: Lateral view of uninjected embryos from 3–24 hpf. D′–H′,D′′–E′′: Lateral view of lzap MO injected embryos. Embryos were time matched to uninjected embryos. Arrows indicate persistence of the syncytial layer. Mild and severe phenotypes following injection of lzap MO are shown.
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