Vav is phosphorylated in a MyD88-dependent fashion and is required for LPS-induced oxidative burst. (A) Oxidative burst in MyD88^−/− and WT PECs stimulated with LPS (10 μg/mL) or PMA (50 ng/mL) was measured by lucigenin chemiluminescence. (B) WT and MyD88^−/− BMDMs were stimulated with LPS (10 μg/mL) for the indicated time points. Cells were lysed immediately following stimulation, and lysates were analyzed by sequential Western blot for phosphotyrosine (pTyr) and Vav. (C) Oxidative burst in Vav^null and WT PECs stimulated with LPS (10 μg/mL) or PMA (50 ng/mL) was measured by lucigenin chemiluminescence. (D-E) Oxidative burst was measured by lucigenin chemiluminescence in BM neutrophils stimulated with (D) PGN (10 μg/mL) or (E) fMLP (10 μM). (F) Oxidative burst in BMDMs stimulated with LPS (10 μg/mL) or PMA (50 ng/mL) was measured with lucigenin. Panels A, C, and F show 2 mice from each genotype that were analyzed within the same experiment. All panels show data representative of more than 5 independent experiments.