Figure 10. The effects of GATA6 expression in EIUM.

(A) Adenoviral-mediated expression of GATA6 (AdGATA6) resulted in significant mRNA (top) and immunoreactive protein (bottom) relative to cells transduced with an adenovirus expressing GFP (AdGFP). IVD did not significantly affect GATA6 expression. (B) qPCR showed that GATA6 transduction was the main affect for the repression of GATA2 and the induction GATA4 mRNA. (C) Immunofluorescence for GATA2 (pseudocolored green), GATA6 (red; middle panels), and ACTA2 (red; right panels) is shown for AdGFP-transduced cells (top row) and AdGATA6-transduced cells (bottom row). Transcription factors showed proper nuclear staining, and GATA2 nuclear signal was reduced when GATA6 was present (DAPI counterstaining is shown beside panels for transcription factors). ACTA2 immunoreactivity was observed in AdGATA6 transduced cells similar to OSIS (compare to figure 1; DAPI counterstain is merged). (D) qPCR was performed on the same panel of 12 genes shown in Figure 1, and EIUM cells transduced with AdGATA6 resulted in a shift in gene expression that resembled OSIS. AdGATA6 was the main effect for the observed change in ESR2, PGR, NR5A1, and CYP19A1. A significant interaction was observed between AdGATA6 treatment and IVD for ESR1, MMP11, FOXO1, HAND2, PRL, and IGFBP. HOXC6 and ALDH1A2 were not significantly affected by either treatment. (n = 4; connecting bars show main effect p<0.05 by 2-way ANOVA; lower case letters indicate significantly different groups p<0.05 by Tukey's after ANOVA).