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. 2007 Feb 16;6(4):664–673. doi: 10.1128/EC.00308-06

FIG. 4.

FIG. 4.

Effect of antisense eh29 on physiological activity and death of the trophozoites during oxidative stress. A: Viability assay of stressed trophozoites using MTT (for details, see Materials and Methods). B: Loss of membrane integrity during oxidative stress. Release of the cytosolic enzyme pyruvate:ferredoxin oxidoreductase served as an indicator of loss of membrane integrity and thus cell viability. Bars: 1, nontransfected trophozoites; 2, pEhHYG-tetR-O-CAT-transfected control trophozoites; 3, uninduced pEhHYG-tetR-O-Reveh29-transfected trophozoites; 4, tetracycline-induced pEhHYG-tetR-O-Reveh29-transfected trophozoites; 5, 5 μM catalase-treated tetracycline-induced transfected trophozoites. Data represent means ± SEM of three independent experiments (P < 0.05). C: The cell death assay was done using an Annexin V-FLUOS kit. The left column represents propidium iodide-positive trophozoites, the middle column represents Annexin-positive trophozoites, and the right column represents a combined image. Panels: 1, nontransfected trophozoites; 2, pEhHYG-tetR-O-CAT-transfected control trophozoites; 3, uninduced pEhHYG-tetR-O-Reveh29-transfected trophozoites; 4, tetracycline-induced pEhHYG-tetR-O-Reveh29-transfected trophozoites; 5, 5 μM catalase-treated tetracycline-induced transfected trophozoites; 6, nontransfected trophozoites exposed to oxidative stress for 6 h.