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. 2014 May;34(10):1812–1826. doi: 10.1128/MCB.01524-13

FIG 6.

FIG 6

Large-scale gene expression profiles in BMMCs transfected with Gata1 siRNA. WT BMMCs were transfected with control or Gata1 siRNA (200 pmol), and total RNA was isolated 24 h after transfection. (A) Pathway analysis was performed using the IPA software program. A comparison of disease-related and biological functions among genes affected in the GATA1 knockdown BMMCs is shown. The red line indicates the threshold for a significant difference (P < 0.05) compared to the BMMCs transfected with control siRNA. (B) The expression levels of the genes listed in the categories of hematological and inflammatory diseases were compared in control and Gata1 siRNA-transfected BMMCs. Heat map comparisons of genes significantly affected (P < 0.05, 2-fold or larger change) in GATA1 siRNA-transfected BMMCs compared to controls are shown. Red indicates relatively high expression, green indicates low expression, and black indicates intermediate expression in GATA1 siRNA-transfected BMMCs compared to controls, as indicated by the color bar. (C) Results of qRT-PCR analysis of three downregulated (Il1a, Epor, and Ahr) and three upregulated (Il18, Tnfrsf11b, and Creg2) genes. The values of the wild-type cells were set to 1.0, and the relative values are shown. (D) Results of qRT-PCR analysis of mast cell-related genes. The values of the wild-type cells were set to 1.0, and the relative values are shown. (E) Configurations of group A tryptase gene loci on mouse chromosome 17A3.3. “Region A” located at the 5′ end of the gene cluster is indicated. (F) Nucleotide sequence of region A. Seven WGATAR sequence motifs are boxed and numbered. All GATA motifs except motif 1 were found to be evolutionally conserved between mouse and rat. The GATA motifs 2, 4, and 6 (in the red box) were identified in recent ChIP-seq studies. (G) The DNA-binding activity of GATA1 to region A, the double GATA region (dblGATA, positive control), and the sixth exon (Gata1 ex6, negative control) of the Gata1 gene was examined using qChIP analysis. The control experiments were performed using rat IgG in place of anti-GATA1 antibodies (N6). *, P < 0.05; **, P < 0.01 (compared to the data for IgG).