Extended Data Fig. 8: METTL3 inhibitor STM2457 affects embryo development and decay of transient genes.
a, Ratio of embryo types at IVF 19 h with and without STM2457 treatment. n = embryo number tested. b, Ratio of embryo types at E4.5 in vitro with DMSO and STM2457 treatment. n = embryo number tested. c, Embryo photos of DMSO and STM2457 treatment at E3.5 and E4.5 in vitro culturing. Scale bar = 100μm. d, m6A marked transcript number in L2C with DMSO and STM2457 treatment. e, Normalized m6A signal intensity in L2C with DMSO and STM2457 treatment. f, RNA signal level of ± 200 bp of stop codon of m6A IP and input for MD m6A + (n = 1089) and ZGA m6A + (n = 756) transcripts defined in Fig. 2a, with DMSO and STM2457 treatment after fertilization. Box plots are presented with horizontal line, median; box, interquartile range (IQR); whiskers, most extreme value within ±1.5 × IQR. P-values are calculated by two-tailed unpaired Wilcoxon test, showed above the boxes. g, PCA of RNA-seq for embryos with DMSO and STM2457 treatment. h, Proportion of m6A- and m6A + transcripts in sustained and transient expressed ZGA transcripts. Two-tailed Fisher’s exact test was used to calculate the p-value. i, Normalized m6A IP and input signal of sustained and transient m6A + transcripts, with DMSO and STM2457 treatment after fertilization. The IP and input signal were scaled by GLuc and CLuc, respectively. j, S/N ratio (targets / Gapdh) in L2C embryos with DMSO and STM2457 treatment after fertilization. Data shown represent 2 biological replicates; value for each biological replicate represents the mean of three technical replicates. k, RT-qPCR results of Zscan4 expression level relative to Actin. l, MERVL RNA level at 4 C and morula relative to 2 C. Data are the mean of MERVL RPKM. m, RT-qPCR results of MERVL RNA level relative to Actin. Data in d-f,i,j represent results from ULI-MeRIP-seq for L2C embryos with two individual replicates. Data in k, m are presented as mean values ± SEM. n = 3 biologically independent samples. P-values are calculated by two-tailed unpair Student’s t test, showed above the bars.
