Fig. 6.

(A) Short-term stimulation of HL60 cells with rGL-7 induces protein tyrosine phosphorylation. After incubation of HL60 cells (2 × 10⁷ cells per ml) for 4 days, 50-μl aliquots were transferred to microtubes, and an equal volume of rGL-7 (20 μg/ml) was added. After 10 s^–10 min, stimulation was terminated by adding 50 μl of SDS sample buffer. For the control, cells were treated with an unrelated Fab antibody (anti-HBs). Protein tyrosine phosphorylation was assessed by immunoblotting using horseradish peroxidase-conjugated PY20. (B) Lyn, Hck, and Cbp coexist with PhGlc in the DIM fraction. Lysates from rGL-7-treated HL60 cells were incubated with Lyn-, Hck-, and Cbp-conjugated protein A agarose gel or biotinylated rGL-7-conjugated avidin agarose gel. The resulting protein complexes were subjected to SDS/PAGE and analyzed by immunoblotting with antibodies against corresponding proteins.