FIGURE 3.

2ME2 induces G₂/M arrest, the displacement of Dab2 from the membrane, and the phosphorylation of the p96 isoform of Dab2. A, semi-confluent ES-2 cells were treated with 2ME2 (4.4 or 10 μm, 16 h) or the same amount of vehicle, suspended by scraping, and their DNA content was measured by propidium iodide staining and fluorescence-activated cell sorting (FACS). B, ES-2 cells, grown on glass coverslips, were treated with 2ME2 (4.4 μm, 16 h), fixed, permeabilized, and stained as indicated in the panels. C, semi-confluent ES-2 cells were treated with 2ME2 (4.4 μm, 16 h), nocodazole (50 μm, 16 h), or vehicle. Cells were lysed, resolved by SDS-PAGE, and immunoblotted with α-Dab2 antibodies D, semi-confluent ES-2 cells were treated for 16 h with 2ME2, lysed prior to immunoprecipitation with α-Dab2 antibodies. Precipitates were treated or not with calf intestinal phosphatase (CIP). Panel depicts a typical experiment (n = 3; the two lanes to the left are the immunoprecipitation (IP), and the two lanes to the right are 10% of the lysates employed).