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Figure 5 | BMC Immunology

Figure 5

From: Proton channel HVCN1 is required for effector functions of mouse eosinophils

Figure 5

Increased membrane depolarization and cytosolic acidification in activated HVCN1-deficient eosinophils. (A) Flow cytometric analysis of eosinophil cell death (7-AAD positive cells) after four-hour PMA (50 ng/ml) stimulation in the presence or absence of ZnCl2 (1 mM) or DPI (10 μM). Data are expressed as the mean ± SD of 3 separate experiments. (B) Mean fluorescence intensity (MFI) of DiBAC4(3) in WT and HVCN1-deficient BM-derived eosinophils. PMA evoked a larger depolarization in HVCN1-deficient than WT eosinophils. The vertical arrow indicates the direction of depolarization. Data are representative of 5 separate experiments. (C) Intracellular pH in WT and HVCN1-deficient BM-derived eosinophils was measured using the pH-sensitive dye SNARF-4 AM (5 μM) at the indicated time points after PMA (50 ng/ml) stimulation. Dead cells were excluded by gating out cells that failed to retain SNARF-4 AM fluorescence. Data are expressed as the mean ± SD of 4 separate experiments. (D) Intracellular pH in WT and HVCN1-deficient BM-derived eosinophils was measured 30 minutes after PMA (50 ng/ml) stimulation in the presence or absence of ZnCl2 at the indicated concentrations. Data are expressed as the mean ± SD of 3 separate experiments. *, P < 0.05; **, P < 0.01; n.s., not significant.

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