Fig. 8.
T5A7-induced Lyn activation in neutrophils.
(A) Neutrophils and DMSO-treated HL-60 cells were incubated in normal IgM–coated (normal IgM) or anti-LacCer IgM T5A7–coated (anti-LacCer IgM) 10 cm dishes at 37°C for indicated periods. Kinase activity was measured by in vitro autophosphorylation as described in “Materials and methods.” Data show the representative time course of 3 independent experiments. To evaluate the recovery of Lyn, the blotted membrane using anti-LacCer IgM T5A7–induced neutrophils was probed with rabbit anti-Lyn IgG (immunoblot). (B) Phosphorylation of Lyn was calculated by scanning the signals of phosphorylated and recovered Lyn by Scion image. The Lyn activity is expressed as fold increase compared with that of cells kept on ice (0 minute). Each point shows the mean ± SD of 3 independent experiments.