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Fig. 3 Identified pure zygotic genes at the RNA level are highly deposited as proteins prior to zygotic genome activation (A) Nested pie charts representing the percentage (%) of mRNAs (internal pie chart) or proteins (external pie chart) classified in the three classes: maternal (red), maternal-zygotic (purple), and zygotic (light blue). (B) Boxplot represents log2 mean protein abundance for each class of genes at each time point (see Figure S4B for reference of expected mRNA dynamics of the same genes), and n is the total number of proteins in each classification. (C) Heatmap showing the mRNA levels of zygotic genes in oocyte and sperm (first five columns), as well as protein (green squares) and mRNA levels (blue squares and last four columns) at 0, 2, 4, and 6 h post-fertilization. Data points represent the log2 mean TPMs (for mRNA, dark blue) and log2 mean protein abundance (dark green) for each stage or time point. Gray squares denote proteins that were not identified at those time points. Labels on the left denote possible origins of deposited zygotic protein, with ? oocytes? being only expressed in oocyte RNA sequencing, ?sperm? only expressed in sperm RNA sequencing, ?both? being expressed in either oocyte or sperm RNA sequencing, ?true zygotic? being proteins that follow expected zygotic patterns of expression, and ?none? being genes that were not found in either oocyte or sperm RNA sequencing. (D) Bar plot from Gene Ontology (GO) term (biological process [BP]) enrichment analysis for zygotic genes. Categories were considered significant when adjusted p < 0.05 (Fisher?s exact test).