Characterization of ΔNZF-GATA-1 and ΔAD-PU.1.

(A) Structure of ΔNZF-GATA-1 and ΔAD-PU.1. (B) Coimmunoprecipitation analyses on the binding activities of ΔNZF-GATA-1 to PU.1 and ΔAD-PU.1 to GATA-1. 293T cells were transfected with HA-tagged FL-GATA-1, HA-tagged ΔNZF-GATA-1, Flag-tagged FL-PU.1, and Flag-tagged ΔAD-PU.1 in various combinations as indicated. Formation of the GATA-1–PU.1 complex was examined by coimmunoprecipitation. (C) Effects of ΔNZF-GATA-1 and ΔAD-PU.1 on the DNA-binding activities of PU.1 and GATA-1, respectively. 293T cells were transfected with ΔNZF-GATA-1, FL-GATA-1, ΔAD-PU.1, and FL-PU.1 in various combinations at the doses indicated. Effects of ΔNZF-GATA-1 (upper panel) and ΔAD-PU.1 (lower panel) on the DNA-binding activities of PU.1 and GATA-1 were examined with the probe of MHC and Mα, respectively. B, DNA-binding complex.