Fig 3

Validation of maternal mRNAs up-regulated in MZznf598.

(A) qRT-PCR analysis of maternal NGD target candidate mRNAs in wild-type (blue) and MZznf598 embryos (red) at 6 hpf relative to 2 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. gstm mRNA (miR-430 target) and ddx4 mRNA (stable maternal mRNA) are shown as controls for Znf598-independent maternal mRNAs. (B) Western blotting to detect Myc-tagged Znf598 proteins at 6 hpf. Tubulin (Tub) was detected as a loading control. (C) Time course qRT-PCR analysis of znf236 and znf970 mRNAs in wild-type (blue) and MZznf598 (red) embryos. (D) Schematics of the Znf236 and Znf970 proteins. C2H2-ZF is indicated in orange. (E) qRT-PCR analysis of znf236 mRNA in wild-type and MZznf598 embryos at 6 hpf injected with control MO (blue and red) or translation-blocking MO (green and orange). (F) PAT assay of znf236 mRNA in wild-type and MZznf598 embryos at 6 hpf. The lane labeled A0 shows the 3′ UTR fragment without a poly(A) tail. (G) Quantification of the PAT assay in (F). The average values of the 2 experiments are shown. The graphs represent the average of 3 independent experiments in A, C, and E. The error bars show the standard deviation. The open circles in A and E show each data point. The numerical data underlying this figure can be found in S1 Data. hpf, hours postfertilization; MO, morpholino oligonucleotide; NGD, no-go decay.