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Fig. 4 mpi morphants are rescued with mpi mRNA. Full-length zebrafish mpi mRNA was cloned using degenerate sequencing primers surrounding the ATG site so that no significant overlap with MO sequence would occur. The embryos were injected with either standard control MO, mpi MO, 6×-myc-mpi mRNA or mpi MO + 6×-myc-mpi mRNA. (A) Western blot showing Mpi expression in embryos from 1 to 5 dpf after mpi mRNA injection at t=0. (B) Restoration of Mpi enzyme activity in embryos injected with either mpi mRNA only or co-injected with mpi MO + mRNA (P=0.47 and 0.80 by paired t-test, respectively, as compared with standard control MO). *P=0.047 by paired t-test. Results from three experiments. (C) mpi morphants show phenotypic rescue when co-injected with 6×-myc-mpi mRNA. Scale bar: 50 μm. (D) At 4 dpf, 78% of morphants injected with 6×-myc-mpi mRNA were rescued to normal as compared with 6% normal in mpi MO embryos; * P<0.0001 by Fisher′s exact test. Numbers in parentheses under n values indicate the number of experiments.