mCLEC-2 can recruit and signal via Syk kinase. A, Western blotting of immunoprecipitates from RAW macrophages using phosphorylated (YP) and unphosphorylated peptides (Y), corresponding to the cytoplasmic tail of mCLEC-2 or Dectin-1. B, Western blotting of anti-HA immunoprecipitates from A20 cells expressing HA-tagged mCLEC-2. Cells were either unstimulated (US) or stimulated with pervanadate (S). Blots were probed with anti-phosphotyrosine (αPY) and anti-Syk as indicated. C, IL-2 production following zymosan stimulation of Syk-deficient (Syk-) and Syk-sufficient. (Syk+) B-cells, transduced with the chimeric receptor. The data shown are the mean ±SD and are representative of at least three independent experiments. *, p <0.05 versus C35 Syk- cells (Student’s t test). D, Confocal image demonstrating the recruitment of phospho-Syk (red) by RAW264.7 macrophages expressing the chimeric receptor following stimulation with FITC-zymosan. The nucleus is stained with Hoechst dye (blue).