FIG. 7.

Ligation of SIRPα promotes the JAK- and PI3-K-dependent recruitment of SHP-1 in macrophages. NR8383 cells were preincubated with AG490 (10 μM; 30 min) or LY294002 (40 μM; 30 min), control cells were incubated with vehicle, and then cells were stimulated with anti-SIRPα (10 μg/ml ED9) for 5 min, lysed, and immunoprecipitated (IP) with anti-SIRPα Abs (rabbit anti-SIRPα from PharMingen in panels A and B) or directly lysed in reducing SDS sample buffer (C). Western blots (WB) of these samples were stained with the indicated antibodies recognizing SHP-1, SHP-2, SIRPα (Santa Cruz Ab), PY, or JAK2 as described in Materials and Methods. HC, heavy chain.