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. 2014 Mar 6;10(3):e1004158. doi: 10.1371/journal.pgen.1004158

Figure 11. The effect of silencing GATA6 and expressing GATA2 in OSIS.

Figure 11

(A) Manipulating GATA expression in OSIS was accomplished using adenoviral transduction to express GATA2 (AdGATA2) and siRNAs targeting human GATA6 (xGATA6). Transduction resulted in significant mRNA (top) and immunoreactive GATA2 (bottom) relative to cells transduced with an adenovirus expressing GFP. Compared to control siRNAs, xGATA6 effectively silenced GATA6 mRNA (qPCR; middle), and protein (immunoblot; bottom). IVD did not significantly affect the expression of any of the GATAs examined, but AdGATA2 strongly repressed (A) GATA6 and (B) GATA4. (C) Phase contrast (left), immunofluorescence for GATA2 (red), and DAPI staining (right) shows nuclear GATA2 protein in AdGATA2-transduced cells. (C) qPCR was performed on the same panel of 12 genes shown in Figure 1. AdGATA2 was the main effect for the observed increase in ALDH1A2, while xGATA6 and AdGATA2 both repressed CYP19A1. IVD was the main effect for FOXO1, HAND2, PRL, and IGFBP. The ESR1, ESR2, PGR, NR5A1, HOXC6 and MMP11 were not significantly affected by any of the treatments. (n = 6; 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).