Figure 3. Ythdf2 is required for m⁶A-dependent RNA decay.

a, RT-qPCR validation of the relative abundance of representative Ythdf2 targets increased at 4 h.p.f. in maternal ythdf2^−/− samples compared with wild-type. b, Design of the reporter RNA. m⁶A-modified (GFP-m⁶A) or non-modified GFP mRNAs (GFP-A) were prepared and co-injected with mCherry control mRNA into one-cell stage embryos. c, The in vivo degradation of GFP-m⁶A mRNAs was faster in wild-type embryos but slightly slower in maternal ythdf2^−/− ones than GFP-A mRNAs. mRNA abundance was determined by RT-qPCR and normalized to 0.5 h.p.f. values. d, Comparison of GFP-m⁶A expression levels in wild-type and maternal ythdf2^−/− embryos at the oblong stage of development (stage-matched embryos, 3.8 h.p.f. for wild-type and 4.2 h.p.f. for mutants). The m⁶A-modified GFP reporter RNA produced consistently higher GFP protein levels in maternal ythdf2^−/− embryos versus wild-type embryos. Quantification of the overall phenomena: 61/72 embryos (85%) for maternal ythdf2^−/− and 59/85 (70%) for wild-type in two experiments. Error bars, s.d., n = 3 (technical replicates). P values were determined using two-sided Student’s t-test for two samples with equal variance.