Journal:

Article Title: Adaptor FYB (Fyn-binding protein) regulates integrin-mediated adhesion and mediator release: Differential involvement of the FYB SH3 domain

doi: 10.1073/pnas.191378198

Figure Lengend Snippet: (A) FYB enhances β-hexosaminidase release in adherent and nonadherent cells. FYB or PEBB (vector control) transfectants grown overnight on 96-well plates were preincubated with DNP-specific IgE. The cells were then either left attached to the plate or dissociated from the well by trypsinization and placed in Eppendorf tubes. Cells were stimulated with 100 ng/ml DNP-KLH either on the plate (adherent) or in the tubes (nonadherent). Data are representative of two experiments with two FYB clones and indicate the mean of triplicate wells. (B) FcɛRI induces tyrosine phosphorylation of FYB in adherent and nonadherent cells. RBL-2H3 cells preincubated with DNP-specific IgE were dissociated from the tissue-culture flask and seeded for 20 min on non-tissue-culture-treated plates that had been coated with either BSA (nonadherent) or FN (adherent). Cells were either unstimulated (Ag, −) or stimulated with DNP-KLH (Ag, +) for 10 min. Lysates were immunoprecipitated with anti-FYB antisera and analyzed by immunoblotting with anti-phosphotyrosine or anti-FYB mAb.

Article Snippet: The cells were then either left attached to the plate or dissociated from the well by trypsinization and placed in Eppendorf tubes.

Techniques: Plasmid Preparation, Clone Assay, Immunoprecipitation, Western Blot