FIGURE 10.

Functional studies of MbSrc1 in mammalian cells. A, SYF cells were transfected with wild-type (WT) or Y530F (YF) forms of human c-Src, WT, or Y489F forms of MbSrc1 or empty vector (C). Lysates were analyzed by SDS-PAGE and anti-Tyr(P) Western blotting. The blots were stripped and analyzed with anti c-Src antibody and with anti-FLAG antibody. B, SYF cells were transfected with MbSrc1 and MbCsk alone or in combination. Lysates were analyzed by Western blotting using anti-Tyr(P) and anti-FLAG (for MbSrc1) antibodies. C, immobilized pYEEI peptide was incubated with lysates from untransfected SYF cells (U), cells transfected with MbSrc alone (S), or cells transfected with MbSrc plus MbCsk (S/C) in buffer containing 50 mm Tris (pH 7.5), 250 mm NaCl, 0.1% Triton X-100, 5 mm EDTA, 0.5 mm sodium vanadate, and 1 mm dithiothreitol. After washing, bound MbSrc was visualized by anti-FLAG immunoblotting. Lysates (25 μg) were loaded directly onto the gel in the right-hand lanes.