a Single-cell zebrafish embryos were injected with random MO, MOs targeting g6pd only, and MOs targeting g6pd with human G6PD or human CDH1 cRNA rescue. Whole-cell lysates were prepared at 72 hpf from embryo injection. G6pd activities were determined through the G6pd activity assay, as described in the Materials and Methods section. Results are expressed as mean ± SD (*p < 0.05 compared with the control MO group). b Protein expressions of E-cadherin and β-catenin in the whole-embryo lysates at 72 hpf were analyzed through Western blot. β-Actin was used as the loading control. One representative Western blot example is shown out of three experiments. c Zebrafish embryos were injected with random MO, MOs targeting g6pd only, or MOs targeting g6pd with human G6PD or human CDH1 cRNA rescue and were monitored through microscopy. Gastrulation was initiated in the MO-injected embryos at 6 hpf. The epiboly rate was decreased in the g6pd MO-injected embryos (top panel). Scale bar, 250 μm. The phenotypic examination of g6pd MO at 16 hpf revealed shedding of the cells at the embryo surface (arrowhead in in the bottom panel). Rescue of the phenotype was performed by coinjecting g6pd MOs with human G6PD or human CDH1 cRNA. n, total number of embryos analyzed from three independent experiments. Scale bar, 250 μm

The kinetics of G6pd activity at 24, 48, and 72 h postfertilization (hpf) are shown. (a)The kinetic data of control MO revealed that G6pd activity was increased during zebrafish embryonic development. However, g6pd MO injection reduced G6pd activity. (b) Cardiac edema was observed in the g6pd morphants (as indicated by the arrowhead).

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