Detection of tyrosine phosphorylated proteins associated with the kinases Lyn, Syk and Axl. Proteins (A: Lyn; B: Syk and C: Axl, D: CDCP-1) were immunoprecipitated from Ks and K-rn cell lysates and subjected to phosphotyrosine Western blotting using a mix of 4G10 and pY-100 antibodies (top panels). Differentially tyrosine phosphorylated proteins are indicated by arrows on the right. After stripping, membranes were cut and blotted against Syk (top), Lyn (middle), Axl (bottom panels). Results are from one experiment representative of 5. (E) Syk was immunoprecipitated from Ks and K-rn cell lysates. Immunprecipitates were subjected to Western-blotting using phosphospecific antibodies against residues Y₃₂₃ and Y_525/526, and reprobed for whole Syk. (F) Syk was immunoprecipitated from K-rn cell lysates from untreated, BAY 61-3606–treated (2μM 2 hours) and PP2-treated (20μM 2 hours) K-rn cells. Immunoprecipitates were subjected to Western blotting using phosphospecific antibodies as described in panel E.