Mapping of regions of GATA-2 and PLZF involved in the interaction by GST pull-down experiments.

(A) GST fusion proteins containing the indicated portions of GATA-2 were tested for the ability to bind Flag-tagged PLZF contained in 293T cell nuclear extract programmed with Flag-PLZF expression plasmids. The first and last amino acids of GATA-2 region present in the various GST fusions are indicated, and the abilities of the proteins to bind PLZF are summarized. Western blot analysis of the pull-down materials using anti-Flag antibody is shown on the right (top panel), and Coomassie brilliant blue (CBB) staining is presented in the lower panel to allow assessment of the quality and quantity of the various GST–GATA-2 proteins used. Numbers on the left indicate positions of molecular weight markers in kilodaltons. (B) Reciprocal pull-down analysis in which various GST–PLZF fusion proteins were analyzed for the ability to bind Flag-tagged GATA-2.