Figure 2
Figure 2. Increased adhesion-induced ROS production in ARAP3-deficient neutrophils. Bone marrow–derived neutrophils from tamoxifen- and mock-induced Arap3fl/flERT2Cre+ mice were prepared (A-B), primed with TNFα and GM-CSF or mock primed, and (all) preincubated with luminol as described in “ROS assays.” Cells (5 × 105) were plated into 96-well plates containing fMLF as a soluble stimulus (A-B) or plates that had been coated with polyRGD (C-D), fibrinogen (E-F), or a BSA–anti-BSA immune complex (G-H), and light emission was recorded using a Berthold Microluminat Plus luminometer. Data were recorded in duplicate and at least 3 independent experiments were performed. Data (mean ± range) from a representative experiment performed with cells from tamoxifen- and mock-induced Arap3fl/fERT2Cre+ mice are shown in panels A, C, E, and G. Data shown in panels B, D, F, and H represent accumulated light emission (mean ± SEM) from the pooled performed experiments (n = 3) expressed as a percentage of the response in mock-induced neutrophils of each particular experiment. Raw data were analyzed by 2-way ANOVA with Bonferroni post tests (B,D,F) and by t tests (Mann-Whitney; H).

Increased adhesion-induced ROS production in ARAP3-deficient neutrophils. Bone marrow–derived neutrophils from tamoxifen- and mock-induced Arap3fl/flERT2Cre+ mice were prepared (A-B), primed with TNFα and GM-CSF or mock primed, and (all) preincubated with luminol as described in “ROS assays.” Cells (5 × 105) were plated into 96-well plates containing fMLF as a soluble stimulus (A-B) or plates that had been coated with polyRGD (C-D), fibrinogen (E-F), or a BSA–anti-BSA immune complex (G-H), and light emission was recorded using a Berthold Microluminat Plus luminometer. Data were recorded in duplicate and at least 3 independent experiments were performed. Data (mean ± range) from a representative experiment performed with cells from tamoxifen- and mock-induced Arap3fl/fERT2Cre+ mice are shown in panels A, C, E, and G. Data shown in panels B, D, F, and H represent accumulated light emission (mean ± SEM) from the pooled performed experiments (n = 3) expressed as a percentage of the response in mock-induced neutrophils of each particular experiment. Raw data were analyzed by 2-way ANOVA with Bonferroni post tests (B,D,F) and by t tests (Mann-Whitney; H).

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