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. 2012 Jul 23;287(36):30641–30652. doi: 10.1074/jbc.M112.355271

FIGURE 5.

FIGURE 5.

SATB2 regulates the physical proximity between Gγ- and Aγ-globin genes. A, SATB2 protein is depicted with domains, including a PDZ domain, a MAR-binding domain (MD) and a homeodomain (HD). B and C, co-IP assay in 293T cells depicting (B) the interaction between differently tagged SATB2 proteins and (C) the PDZ domain dependence of the interaction. D, luciferase reporter assay in 293T cells showed that deletion of the PDZ domain of SATB2 abolished the transcriptional activity of SATB2. Each error bar represents a standard deviation calculated from experiments performed in triplicate. Student's t test was used for statistical analysis, and p value is indicated (**, p < 0.01). E, 3C assay of HindIII fragments covering the indicated regulatory elements of the β-globin locus in K562 cells. Perturbation of SATB2 expression specifically affected the physical proximity between the Gγ- and Aγ-globin promoters. Each error bar represents a standard deviation calculated from experiments performed in triplicate. Student's t test was used for statistical analysis, and p values are indicated (*, p < 0.05). F, ChIP-3C assay with anti-Myc antibody in K562/SATB2-myc cells confirmed that SATB2 mediated the physical association of the Gγ- and Aγ-globin promoters.