Figure 2
Figure 2. Annexin V–depleted BM neutrophils from G6pc3−/− mice exhibit impaired function. The annexin V–depleted BM neutrophils were isolated from 6- to 8-week-old unaffected (+/+, ○) and G6pc3−/− (−/−, ●) littermates as described in “Depletion of apoptotic cells from neutrophils.” (A) Neutrophil viability. The viability of annexin V–depleted BM neutrophils before (Glucose −) or after incubation for 30 minutes in 5.6mM glucose (Glucose +) was estimated by trypan blue exclusion. Results represent the mean ± SEM of quadruplet determinations. (B) Neutrophil respiratory burst activity in response to 200 ng/mL PMA. (C) Neutrophil concentration–dependent chemotaxis in response to fMLP and KC. *P < .05. (D) Calcium flux in response to 10−6M fMLP or KC. Representative experiments are shown. (E) Neutrophil phagocytosis activity. Representative immunofluorescence of cells with phagocytosed pHrodo E coli bioparticles (red fluorescence) and DAPI nuclei staining (blue fluorescence) at 400× magnification, and quantification of bioparticle-positive neutrophils incontrol and G6pc3−/− mice. Data represent the mean ± SEM of 3 independent experiments. **P < .005.

Annexin V–depleted BM neutrophils from G6pc3−/− mice exhibit impaired function. The annexin V–depleted BM neutrophils were isolated from 6- to 8-week-old unaffected (+/+, ○) and G6pc3−/− (−/−, ●) littermates as described in “Depletion of apoptotic cells from neutrophils.” (A) Neutrophil viability. The viability of annexin V–depleted BM neutrophils before (Glucose −) or after incubation for 30 minutes in 5.6mM glucose (Glucose +) was estimated by trypan blue exclusion. Results represent the mean ± SEM of quadruplet determinations. (B) Neutrophil respiratory burst activity in response to 200 ng/mL PMA. (C) Neutrophil concentration–dependent chemotaxis in response to fMLP and KC. *P < .05. (D) Calcium flux in response to 10−6M fMLP or KC. Representative experiments are shown. (E) Neutrophil phagocytosis activity. Representative immunofluorescence of cells with phagocytosed pHrodo E coli bioparticles (red fluorescence) and DAPI nuclei staining (blue fluorescence) at 400× magnification, and quantification of bioparticle-positive neutrophils incontrol and G6pc3−/− mice. Data represent the mean ± SEM of 3 independent experiments. **P < .005.

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