FIG. 7.

Effects of PI 3-kinase inhibition on Stx7 SNARE complexes in macrophages. Mouse bone marrow-derived macrophages were deprived of CSF-1 for 16 h before being treated with 0.1% dimethyl sulfoxide or 10 μM LY294002 for 30 min. The macrophages were stimulated with CSF-1 for 15 min and then lysed. Vti1b (A and B), Stx8 (C and D), and Stx7 (E) were subsequently immunoprecipitated (IP) from the cell lysates with anti-Vti1b (α-Vti1b), anti-Stx8 (α-Stx8), and anti-Stx7 (α-Stx7) antibodies, respectively, and then Western blotted with the indicated antibodies. (B and D) Quantified data are presented as the increase in SNARE protein binding following CSF-1 stimulation relative to that in unstimulated macrophages. Data represent the means (± standard errors) of three experiments. (F) Western blotting of cell lysates with the indicated antibodies.