Figure 2.
Cell motility, CD11b expression, and F-actin content and assembly. (A) Chemotaxis measured using the under-agarose technique showing distance migrated for each stimulus (mean ± SEM) for the control and patient’s neutrophils, respectively, in 3 to 4 experiments, using independent neutrophil isolation. *P < .05, by 2-tailed Student t test,. (B) Chemotaxis measured using the transmembrane migration technique in response to fMLF. Fluorescence-labeled patient cells and control cell were detected as they crossed the fluorescence-blocking PET membrane into the lower wells in the system. Fluorescence moving into the lower wells is expressed as a percentage of the fluorescence detected in 106 labeled cells and is plotted over time. Directed migration was decreased in the patient’s cells; nondirected migration with buffer was also decreased in the patient’s cells (data not shown). (C) Flow cytometry histogram showing a representative example of CD11b expression for control and patient neutrophils after incubation with buffer, PMA, and fMLF. (D) Cell surface levels of CD11b determined by flow cytometry and presented as mean log fluorescence after incubation with buffer, PMA, and fMLF for the control (red) and patient’s (blue) neutrophils. Bars and brackets represent the mean ± SEM of results in 3 to 5 experiments on independent neutrophil isolations. (E) F-actin assembly was measured as in “Methods” and plotted as mean log fluorescence for buffer, PMA, and fMLF for the control (red) and the patient (blue). The bars and brackets are ±SEM of results in 3 experiments using independent neutrophil isolations. (D-E) *P < .05, patient vs control in a given treatment condition, and #P < .05, given stimulus vs buffer, by 2-tailed Student t test.

Cell motility, CD11b expression, and F-actin content and assembly. (A) Chemotaxis measured using the under-agarose technique showing distance migrated for each stimulus (mean ± SEM) for the control and patient’s neutrophils, respectively, in 3 to 4 experiments, using independent neutrophil isolation. *P < .05, by 2-tailed Student t test,. (B) Chemotaxis measured using the transmembrane migration technique in response to fMLF. Fluorescence-labeled patient cells and control cell were detected as they crossed the fluorescence-blocking PET membrane into the lower wells in the system. Fluorescence moving into the lower wells is expressed as a percentage of the fluorescence detected in 106 labeled cells and is plotted over time. Directed migration was decreased in the patient’s cells; nondirected migration with buffer was also decreased in the patient’s cells (data not shown). (C) Flow cytometry histogram showing a representative example of CD11b expression for control and patient neutrophils after incubation with buffer, PMA, and fMLF. (D) Cell surface levels of CD11b determined by flow cytometry and presented as mean log fluorescence after incubation with buffer, PMA, and fMLF for the control (red) and patient’s (blue) neutrophils. Bars and brackets represent the mean ± SEM of results in 3 to 5 experiments on independent neutrophil isolations. (E) F-actin assembly was measured as in “Methods” and plotted as mean log fluorescence for buffer, PMA, and fMLF for the control (red) and the patient (blue). The bars and brackets are ±SEM of results in 3 experiments using independent neutrophil isolations. (D-E) *P < .05, patient vs control in a given treatment condition, and #P < .05, given stimulus vs buffer, by 2-tailed Student t test.

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