Figure 5
Cell lysis of H. pylori induced by the action of VDP1.
(a) FC-retaining H. pylori was incubated for 48 h in the presence or absence of VDP1 at a 10 μg/ml concentration in PBS containing dMβCD (30 μM) under an anaerobic condition and the CFUs were counted. Three independent experiments were carried out to calculate the mean CFU ± SD per ml. (b) The OD660 nm of bacterial suspensions was measured after performing the same experiment described in panel (a). The statistical significance of differences in OD660 nm between the bacterial suspensions in the presence and absence of VDP1 was evaluated by the t-test based on data obtained from three independent pair-experiments. (c) H. pylori cells obtained from the experiment described in panel (a) were microscopically observed. (d) The cell membrane lipids of FC-retaining H. pylori were extracted from the bacterial cell supernatant obtained from the same experiment described in panel (a), in order to quantify the FC amounts via the ferrous chloride-sulfuric acid method. The statistical significance of differences in FC amounts detected in the bacterial cell supernatants in the presence and absence of VDP1 was evaluated by the t-test based on data obtained from three independent pair-experiments. The “0 h” in panels (a), (b), (c) and (d) indicates the CFU, OD660 nm, bacterial shape and FC amount analyzed immediately before exposure to the anaerobic atmosphere, respectively.
