Fig. 3.
Far Western analysis using metabolically labeled S35-GST-Grb4.
We precipitated 5 mg of K562 RIPA protein extract at 4°C overnight with 10 μg of GST or equimolar amounts of various GST fusion proteins as indicated. Bound proteins were resolved on SDS-PAGE before being blotted, blocked, and renatured at room temperature. The membrane was probed with S35-GST-Grb4 and analyzed by autoradiography. Solid arrows indicate proteins bound strongly by GST-Grb4 and either weakly or not at all by Gst-Nck; open arrows indicate proteins that bind both to Gst-Grb4 and to Nck but show marginally stronger binding to Grb4. The solid circle indicates GST-Nck.
