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. 2025 Sep 23;53(18):gkaf956. doi: 10.1093/nar/gkaf956

Figure 4.

Figure 4.

Function of Eif1ad3 for mouse embryo development. (A) Heatmap showing the expression level of Eif1 family genes. The color spectrum, ranging from red through white to blue, indicates high to low levels of gene expression. (B) Multiple sequence alignment (Omega) of EIF1AD amino acid sequence. (C) Experimental scheme of Eif1ad3 knock down experiment. (D) A representative image of mouse embryos from control and KD groups. (E) Four-cell developmental rate of Eif1ad3 KD embryos compared to control group. (F) Volcano plots showing the number of upregulated or downregulated genes in control compared to Eif1ad3 KD at two-cell stage, the top GO terms of upregulated and downregulated genes are shown respectively. (G) Venn diagram showing the number of genes and the top representative GO terms that are overlapped between control enriched and two-cell specific genes from HF profile in Fig. 2A. (H) Venn diagram showing the number of genes and the top representative GO terms that are overlapped between control enriched and four-cell specific genes from HF profiles in Fig. 2A. (I) Rank the DEGs according to the log2 fold change. Each circle represents a gene, and the highlighted circle represents the Rpl and Eif genes separately. The genes are annotated with detailed information listed in Supplementary Dataset 5. (J) Representative confocal images of two-cell embryos with incorporation of methionine analog (HPG). From three biological replicates, n ≥ 21; HPG, gray and red; H2B-GFP, green; 4′,6-diamidino-2-phenylindole (DAPI), blue; scale bar, 20 μm. NTC, negative technical control where HPG was omitted. (K) Quantification of HPG signal from panel (J). Values are presented as mean ± SD; Student’s t-test: **P <.01. (L) Analysis of global de novo proteosynthesis by incorporation of 35S-methionine in embryos with downregulation of Eif1ad3. Data from four biological replicates. GAPDH was used as a loading control. (M) Quantification of 35S-methionine incorporation from panel (L). Values from control KD were set as 100%. Data presented as mean ± SD; Student’s t-test: *P <.01.