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Fig. 2 | Parasites & Vectors

Fig. 2

From: Trichomonas vaginalis induces apoptosis via ROS and ER stress response through ER–mitochondria crosstalk in SiHa cells

Fig. 2

Trichomonas vaginalis induced mitochondrial apoptosis in SiHa cells. SiHa cells were infected with live T. vaginalis trophozoites at MOI 2 and 5 for 0, 2 or 6 h. a PARP and caspase-3 proteins were assessed using western blotting analysis, and anti-β-actin was used as a loading control. Lanes: Tv only, T. vaginalis parasite protein without SiHa cells; M, protein marker; CTL, untreated control SiHa cells; Tv, SiHa cells infected with live T. vaginalis. b Ratio of apoptosis was measured and analyzed by Annexin V-FITC/PI staining and flow cytometry. Cells that are considered to be viable are both FITC Annexin V and propidium iodide (PI) negative; cells that are necrotic cells are FITC Annexin V negative and PI positive; cells that are in early apoptosis are FITC Annexin V positive and PI negative; and cells that are in late apoptosis or already dead are are both FITC Annexin V and PI positive. Data are presented as the means ± SD, and asterisks indicate significant difference (***P < 0.001) compared to the control group. c SiHa cells were pretreated with various concentrations of an ROS scavenger (NAC) for 2 h, and subsequently infected with T. vaginalis at MOI 5 for 6 h. PARP and cleaved caspase 3 protein levels were assessed by western blotting. d JC-1 staining was observed by confocal fluorescence microscopy. In JC-1-stained cells, red fluorescence is visible in cells with high mitochondrial membrane potential, while green fluorescence of JC-1 monomer is present in cells with low mitochondrial potential. e SiHa cells were pretreated with various concentration of NAC for 2 h and subsequently infected with T. vaginalis at MOI 5 for 6 h. JC-1 staining was observed using confocal fluorescence microscopy. Scale bars: 10 µm. Data shown are representative of three independent experiments with similar results

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