Fig 4

Fig 4 Validation of zygotic C2H2-ZF mRNAs up-regulated in MZznf598.

(A) Cumulative distributions of fold changes in mRNA levels in MZznf598 embryos compared to wild-type embryos at 6 hpf. C2H2-ZF genes on chromosome 4 (orange), C2H2-ZF genes on other chromosomes (purple), and genes without C2H2-ZF (black) are shown. The x-axis shows the fold change, and the y-axis shows the cumulative fraction. The p values are shown on the left (Kolmogorov–Smirnov test). (B) qRT-PCR analysis of C2H2-ZF mRNAs in wild-type (blue) and MZznf598 embryos (red) at 6 hpf. The results for MZznf598 embryos rescued by injecting mRNAs encoding Myc-tagged full-length Znf598 (light blue) or a mutant Znf598 lacking the RING domain (pink) are also shown. actb1 mRNA is shown as a control for Znf598-independent mRNA. The chromosomal locations are indicated below the gene name. (C) A scheme of typical FZNF mRNA and corresponding pre-mRNA/genetic loci. (D) Genomic qPCR analysis using primers that amplify exons encoding the FiNZ domain. The estimated copy number of FiNZ exons per haploid genome normalized to that of rpl24 is shown on the y-axis. (E) qRT-PCR analysis of FZNF mRNAs and pre-mRNAs at 6 hpf. The colors are the same as in (B). (F) qRT-PCR analysis of FZNF mRNAs at 6 hpf in wild-type embryos injected with control MO (blue), znfMO4 (light green), or FiNZ ATGMO (green). znf292b (a C2H2-ZF gene without the FiNZ domain) and actb1 (a gene without C2H2-ZF) are shown as controls. The graphs in B, E, and F represent the average of 3 independent experiments. The graphs in D represent the average of 8 independent embryos. The error bars show the standard deviation. The open circles in B, D, E, and F show each data point. Asterisks in E and F indicate p < 0.05 (Dunnett’s test, compared to wild type or control MO). The numerical data underlying this figure can be found in S1 Data. hpf, hours postfertilization; MO, morpholino oligonucleotide.