Skip to main content
. 2013 Nov 6;123(3):366–376. doi: 10.1182/blood-2013-05-500207

Figure 2.

Figure 2

Neutrophils isolated from patients with gain-of-function HIF2A mutations have enhanced survival but normal function. (A) Apoptosis. Neutrophils from patients with HIF2A mutations (closed squares) or healthy controls (open squares) were cultured for 20 hours with dimethyloxalylglycine (0-100 µM) and apoptosis was determined by morphology. Data are mean ± SEM for n = 3, analyzed by 2-way ANOVA with Bonferroni’s multiple comparison post-test. Overall, there was a significant difference between the control group and the patient group (P < .001), with multiple comparison testing showing statistical significance at the 1 µM concentration (*P < .05). (B) Expression of the HIF targets VEGF, PAI1, and PHD3. TaqMan quantitative PCR analysis of cDNA prepared from freshly isolated neutrophils of patients with HIF2A mutations (filled bars) or healthy controls (open bars). Data are mean and SEM for n = 3. (C-D) Phagocytosis. (C) Phagocytic index was calculated from cytospin slides of neutrophils from healthy controls (open bars) and HIF2A patients (filled bars) prepared after 30 minutes of culture with opsonized zymosan (0.2-1 mg/mL). (D) Flow cytometry analysis of intracellular Alexa Fluor 488 E coli was performed after 30 minutes of culture of cells from healthy controls (open bars) and HIF2A patients (filled bars). (E) Respiratory burst. Neutrophils from healthy controls (open bars) and HIF2A patients (filled bars) were cultured in the presence of dichlorofluorescin only or dichlorofluorescin and N-formyl-Met-Leu-Phe (100 nM) or zymosan A (0.2 mg/ml) and analyzed by flow cytometry. Data show mean and SEM for n = 3.