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. 2013 Mar 15;8(3):e58828. doi: 10.1371/journal.pone.0058828

Figure 5. Changes in yolk sac gene expression in cKO embryos.

Figure 5

A–E) RT-PCR and qPCR performed with cDNA prepared from whole 9.0 and 9.5 dpc cKO and WT yolk sacs. A) As expected, Yy1 is significantly downregulated in whole cKO yolk sacs. β-actin and Hprt expression are used as loading controls. B) No expression differences between WT and cKO samples are noted for VegfA using primers that recognize all (Exon 2–3) or the alternative VegfA isoforms (Exons 3–8) nor in the Vegf transcriptional regulator Hif1α. C) While many visceral endoderm-specific genes show no expression differences, expression of vHnf1, Hnf4α and Pgc1α were all downregulated in cKO samples when compared to WT at 9.0 and 9.5 dpc. D) An examination of genes involved in lysosome biogenesis reveals no expression differences between WT and cKO yolk sacs with the exception of Enpp-2, which is upregulated in mutant samples at both stages examined. E) qPCR reveals that Yy1 is expressed at ∼30% of WT levels in whole yolk sacs, where mesoderm derivatives maintain Yy1. qPCR was used to confirm that VegfA expression is not significantly altered between cKO and WT and that expression of the visceral endoderm gene, Hnf4α is significantly downregulated in cKO yolk sacs. *** = p value<0.001; error bars = standard error.