Fig 6

Analysis of the effect of C2H2-ZF sequences on mRNA stability.

(A) A scheme of the mRNA injection assay used to measure the effect of the sequence to be tested (shown as X in gray) on mRNA stability. (B) qRT-PCR analysis of injected sfGFP reporter mRNAs at 6 hpf relative to 2 hpf in wild-type (blue) and MZznf598 (red) embryos. (C) qRT-PCR analysis of injected sfGFP-ZF6×3 reporter mRNAs at 6 hpf relative to 2 hpf in wild-type (blue), MZznf598 (red), GFP MO-injected wild-type (light green), and GFP MO-injected MZznf598 (orange) embryos. The graphs in B and C represent the average of 3 independent experiments. The error bars show the standard deviation. The open circles show each data point. Asterisks indicate p < 0.05 (Student’s t test in B and Dunnett’s test compared to the wild type in C). (D) A scheme of the disome localization score analysis. (E) Cumulative distributions of fold changes in mRNA levels in MZznf598 embryos compared to wild-type embryos at 6 hpf. All genes (black) and C2H2-ZF genes with high (dark blue), middle (blue), and low (turquoise) disome localization scores are shown. The x-axis shows the fold change, and the y-axis shows the cumulative fraction. The p values are shown on the right (Kolmogorov–Smirnov test). (F) A model of NGD for degrading mRNAs encoding stall-prone tandem C2H2-ZF sequences in zebrafish. The numerical data underlying this figure can be found in S1 Data. hpf, hours postfertilization; MO, morpholino oligonucleotide; NGD, no-go decay.