Mi2β Is Required for γ-Globin Gene Silencing: Temporal Assembly of a GATA-1-FOG-1-Mi2 Repressor Complex in β-YAC Transgenic Mice
Figure 6
Human β-like globin expression in adult −566 Aγ-globin HPFH β-YAC transgenic mice.
(A and B) Expression of the fetal γ- and adult β-globin was analyzed by real-time qRT-PCR and the data was normalized to mouse α-globin or Gapdh gene expression. (A) Human fetal γ-globin mRNA expression; B) Adult β-globin mRNA expression. Lines are indicated at the bottom of the plot. Results are the average of triplicate experiments from three animals of lines 20. * indicates P<0.05. (C–F) Flow cytometry analysis was performed using a mouse monoclonal anti-γ-globin antibody to determine the percentage of F cells. C) Wild-type β-YAC; D) −117 Greek HPFH β-YAC; E) −566 Aγ-globin HPFH β-YAC transgenic line 20 (1), F) −566 Aγ-globin HPFH β-YAC transgenic line 20 (2). Flow cytometry controls are identical to those shown in Figure 2A, 2B and 2G since the experiments were performed together. (G–I) Cytospins were prepared using an anti-human hemoglobin F FITC-conjugated antibody. G) Wild-type β-YAC; H) −117 Greek HPFH β-YAC; I) −566 Aγ-globin HPFH β-YAC transgenic line 20. Top panels, visible light images of cytospin cells; bottom panels, immunofluorescent images of same fields.