Fig. 7.
Both tyrosine residues in FcγRIIa ITAM are responsible for SHIP phosphorylation in vivo.
(A) The expression of CD8 chimeras with substitutions of tyrosine residues with phenylalanine was examined by FACS analysis using biotinylated F(ab′)2 fragments of OKT8 followed by FITC-conjugated streptavidin. Dotted lines indicate fluorescence of unstained cells. (B) The THP-1 transfectants were stimulated with biotinylated F(ab′)2 fragments of OKT8 followed by streptavidin. The lysates from the cells were immunoprecipitated with anti–SHIP antibody, separated on SDS-PAGE gels, and blotted with antiphosphotyrosine (anti-pTyr) antibody. The filter was reprobed with anti–SHIP antibody.
