GATA-2 interacts with HDAC5 in vivo and in vitro.

(A) pcDNA3.1/His–GATA-2 (5 μg) was cotransfected with 5 μg of either Flag-HDAC4, Flag-HDAC5, or Flag-HDAC6 into COS cells. Whole cell lysates were prepared and immunoprecipitated with anti–X-press antibody. Immunoprecipitates were subjected to immunoblot analysis with anti-Flag antibody (upper panel). The blot was reprobed with anti–X-press 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 class II HDAC was highly expressed. (B) GATA-2 interacts with HDAC3 and 5 in vitro. [³⁵S]-methionine-labeled HDAC1, HDAC3, and HDAC5 translated in vitro were incubated with GST–GATA-2 fusion protein or GST protein alone and pulled down with GST-agarose beads. After 4 washes, the precipitates were dissolved by SDS-PAGE and visualized by autoradiography. Input represents 4% of in vitro–translated HDAC proteins used for the pull-down assay.