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Figure 2 | BMC Immunology

Figure 2

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

Figure 2

ROS production from WT and HVCN1-deficient eosinophils. (A) Time-dependent H2O2 production by WT and HVCN1-deficient (KO) BM-derived eosinophils was measured using HBSS solution containing 25 μM Amplex Red and 0.05 U/ml HRP. Eosinophils were stimulated by PMA (50 ng/ml) in the presence or absence of Zn2+ or DPI at the indicated concentrations. Data are representative of 4 separate experiments and expressed as mean ± SD. The arrow indicates the addition of PMA at Time = 0. (B) Mean H2O2 production under the same conditions as (A) at 60 minutes following PMA stimulation. Data are expressed as the mean ± SD of 4 separate experiments done in triplicate. (C) Time-dependent superoxide production by WT and HVCN1-deficient BM-derived eosinophils was determined using HBSS solution containing lucigenin (200 μM). Eosinophils were stimulated by PMA (50 ng/ml) in the presence or absence of SOD (1000 U/ml). Data are representative of 4 separate experiments and expressed as mean ± SD. The arrow indicates the addition of PMA at Time = 0. (D) Mean superoxide production under the same conditions as (C) at 30 minutes after PMA stimulation. Data are expressed as the mean ± SD of 4 separate experiments done in triplicate. *, P < 0.05; **, P < 0.01; n.s., not significant.

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