FIG. 2.

Interaction of Dvl-1 with Dvl-3. (A) Intact cells. Lysates (20 μg of protein) of COS cells expressing Myc–Dvl-3 (lane 2), HA–Dvl-1 (lane 3), HA–Dvl-1-(140-670) (lane 4), Myc–Dvl-3 and HA–Dvl-1 (lane 5), or Myc–Dvl-3 and HA–Dvl-1-(140-670) (lane 6) were simultaneously probed with anti-Myc and anti-HA antibodies. The same lysates (200 μg of protein) were immunoprecipitated with the anti-Myc antibody, and the immunoprecipitates were probed with the anti-Myc and anti-HA antibodies (lanes 7 to 11). The lysates of COS cells transfected with empty vectors were used as a control (lane 1). IP, immunoprecipitation; Ab, antibody; Ig, immunoglobulin. The arrows and arrowhead indicate the positions of HA–Dvl-1 or HA–Dvl-1-(140-670) and Myc–Dvl-3, respectively. (B) Direct binding. After GST–Dvl-1-(1-82) (lanes 1 to 3) or GST–Dvl-1-(140-670) (lanes 4 to 6) (50 pmol of each) was incubated with MBP–Dvl-3 (full length) (lanes 1 and 4), MBP–Dvl-3-(1-80) (lanes 2 and 5), and MBP (lanes 3 and 6) (30 pmol of each) immobilized on amylose resin, MBP fusion proteins were precipitated by centrifugation. The precipitates were probed with the anti-GST antibody. The positions of GST–Dvl-1-(1-82) and GST–Dvl-1-(140-670) are shown in lanes 7 and 8, respectively. The results shown are representative of three independent experiments.