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Figure 5 Mechanistic target of rapamycin (mTOR) signaling pathway disrupted in 24 hpf E– embryos. (A) E+ and E– embryos at 24 hpf metabolomic data acquired previously were integrated with gene expression profiles using MetaboAnalyst to generate network profiles. The mTOR signaling pathway was significantly enriched with both metabolites and genes expressed in 24 hpf E– embryos. Red boxes or circles represent increased, while green boxes represent reduced metabolite or gene expression, respectively, in E– relative to E+ embryos. (B) mTOR complex 1 (mTORC1)-associated proteins including Raptor, Rps6k, Eif2a and Eif4ebp1 were evaluated by activity determined by phosphorylation status in pooled protein extracts of rat liver (positive control), E+ and E– embryos (n = 30 embryos/pool) at 24 hpf. Phospho-protein (p-) to unphosphorylated protein ratio is calculated for the representative lanes shown.