Effects of EphB4 overexpression on cell lines.

(A) CMK cells were transduced with MSCV-GFP (left) or MSCV-EphB4 (right). GFP⁺ cells were sorted and cultured in the absence or the presence of 100 nM PMA for 24 hours. Expression of the megakaryocytic marker CD41 was evaluated by FACS using anti-CD41 PE-conjugated monoclonal antibody. Data from 3 independent experiments are summarized in the text. (B) Ploidy analysis of CMK and CMY. CMK or CMY cells transduced by MSCV-GFP (left) or MSCV-EphB4 (right) were sorted and cultured for 3 days. Cells were fixed and stained with 50 μg/mL propidium iodide. Ploidy values were determined by plotting propidium iodide fluorescence of the cells using a semilogarithmic scale. (C) HL-60 cells transduced with MSCV-GFP (left) or MSCV-EphB4 (right). GFP⁺ cells were sorted and cultured in the presence of 2 μM all-trans retinoic acid for 5 days (upper) or in the presence of 4 nM PMA for 3 days (lower). Other time point data were not shown here. Expression of the granulocytic and monocytic marker CD11b was evaluated by FACS using anti-CD11b PE-conjugated monoclonal antibody. Data presented are from a representative experiment of 3 performed for panels A, B, and C.