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

FIGURE 2.

FIGURE 2.

SATB2 activates human γ-globin gene transcription in erythroid cells. A and B, retroviral vectors for (A) SATB2 overexpression or (B) SATB2 knockdown were transfected into human primary erythroid cells at day 1 of differentiation. The expression of SATB2 and β-like globin genes was determined on day 4 using Western blot and real-time RT-PCR. 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 < 0.01). C and D, K562 cells were infected with a retrovirus harboring (C) Myc-tagged SATB2 cDNA or (D) short hairpin RNA to knockdown SATB2. Western blot analysis of SATB2, SATB1 and important erythroid transcription factors and real-time RT-PCR analysis of β-like globin genes were performed. 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 < 0.01). E and F, quantitative ChIP analysis of (E) histone modifications and (F) RNA polymerase II presence at the γ-globin promoter in K562 cells after SATB2 overexpression or knockdown. 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).