Effect of GATA-2 on the expression and function of cell cycle regulatory molecules.

(A) The cells of Ba/F3/GATA-2/ER and FDC-P1/GATA-2/ER were cultured with mIL-3 in the presence of 1 μM estradiol and subjected to Northern blot analysis at the time indicated. Similar results were observed in 2 other clones of GATA-2/ER–transfected Ba/F3 and FDC-P1. (B,C) The expression levels of p21^WAF1 and p27^Kip1 proteins were examined by Western blot analysis on total cellular lysates. CDK2 was immunoprecipitated from equal amounts of the cell lysates. Immune complex kinase assays were performed in kinase buffer containing 5 μg Histone H1 and 20 μCi (0.74 MBq) [γ-³²P]ATP for 30 minutes at 30°C. Then, samples were subjected to SDS-PAGE. The gels were stained with Coomassie blue, destained, dried, and subjected to autoradiography. The results shown in panels B and C are representative of 3 independent experiments.