Fig. 6: Cxxc1 and Kmt2b are required for proper ZGA and embryonic development.

a Illustration of experimental setup. b (Left) Western blot showing H3K4me3 levels in Cxxc1 and Kmt2b knockdown embryos compared to non-injected control and control morpholino-injected embryos at pre-ZGA. (Right) Quantification of H3K4me3 signal intensity relative to H4 for each condition for three independent experiments. Data are presented as mean values ± SD. c Volcano plots displaying expression log2 fold change and adj. p-value (cutoff: 0.05) in knockdown conditions compared to control morpholino for biological replicate 1 at 7.5 hpf, colored by RNA dynamics groups. d Expression of zygotic genes in 3 technical replicates of every sample for biological replicate 1, clustered by H3K4me3 dynamics group (SHARED, GAINED and ABSENT). Log2 fold change of every gene is calculated over the mean TPM of three technical replicates of the control morpholino of the respective time point. Genes are annotated with differential expression status separately for the Cxxc1 and Kmt2b knockdown conditions, as well as by RNA dynamics group. e Log2 fold change of gene expression levels of all zygotic genes calculated over mean TPM of control morpholino technical replicates for biological replicate 1 for SHARED (top, n = 473) and GAINED (bottom, n = 121) groups at 7.5 hpf. Statistical test: one-sided Wilcoxon rank-sum test with alternative hypothesis that values of left distribution tend to be larger than right distribution; p: (****) ≤ 0.0001, (***) ≤ 0.001, (**)≤0.01, (*) ≤ 0.05; n.s. are p > 0.05. Box plots show the median (center line), 25–75th percentiles (box), whiskers extending to 1.5× the interquartile range, and outliers as individual points. f Log2 f old change expression of pou5f3.2 and sox3 calculated over mean TPM of control morpholino replicates at 7.5 hpf (n = 12). Both biological replicates are represented, each by a different point style. Error bars indicate mean ± SD. g Representative phenotype images of embryos for each condition at NF41. n = 2 experiments, N > 35 per condition. h Quantification of survival phenotype. n = 2 experiments, N > 35 embryos per condition. Source data are provided as a Source Data file.