GATA-2 interacts with HDAC3 in vivo.

(A) Flag–GATA-2 (5 μg) was cotransfected with Myc-HDAC1 (5 μg) or Myc-HDAC3 (5 μg) into COS cells. Whole cell lysates were prepared and immunoprecipitated with anti–Flag M2 beads. Immunoprecipitates were subjected to immunoblot analysis with anti-Myc antibody (upper panel). The blot was reprobed with anti-Flag antibody to confirm that GATA-2 was successfully immunoprecipitated (lower panel). Input represents 5% of whole cell lysates used for each immunoprecipitation applied to confirm whether each HDAC or GATA-2 was highly expressed. (B) The same lysates described above were immunoprecipitated with anti-Myc antibody followed by immunoblot analysis with anti-Flag antibody (upper panel). Successful immunoprecipitation was confirmed by reprobing with anti-Myc antibody.