Journal: PLOS Neglected Tropical Diseases

Article Title: Distinct features of the host-parasite interactions between nonadherent and adherent Trichomonas vaginalis isolates

doi: 10.1371/journal.pntd.0011016

Figure Lengend Snippet: A. TH17 trophozoites (a) were co-incubated with h VECs for 15 (b. and c.), 30 (d, e, f), or 60 min (g and h) for SEM. White triangles point to the flagellum (Fg), axostyle (Ax), or membrane protrusions (Pt). The representative morphology was observed from at least ~50 trophozoites at stages as specified in the infection course. B. The trophozoites treated with DMSO or CytD were cultured in T25 flasks for 1 hr and the morphology was monitored by phase-contrast microscopy. The black and white arrowheads respectively indicate the amoeboid and flagellate forms of T . vaginalis . The percentage of flagellate against amoeboid trophozoites was measured in 600 trophozoites within 6 microscopic fields as shown in the bar graph. C. TH17 trophozoites treated with DMSO or CytD were fractionated into F-actin and G-actin-containing fractions for western blotting. The ratio of the α-actin signal in G-actin versus F-actin was quantified as shown in the bar graph. α-Tubulin (α-Tub) and Tv CyP2 were detected as the purity markers of F-actin and G-actin fractions, respectively. D. The trophozoites pretreated with DMSO or CytD were co-cultured with h VECs (moi 1:3) for 1 hr in cytoadherence binding assay. The number of trophozoites bound per 1000 h VECs was measured as shown in the bar graph (mean ± SD). All experiments were repeated three times. Significant differences were statistically analyzed by the Student’s t-tests, with P <0.01(**) and P <0.05(*).

Article Snippet: The membrane was sequentially incubated with mouse anti-α-actin (10,000× Ac-40, Genetex), mouse anti-α-tubulin (10,000×, Sigma, DM1A), mouse anti- Tv CyP2 (3,000×) [ ], and mouse anti-GAPDH (5,000×, a gift from Dr. John Alderete, Washington State University) and horseradish peroxidase (HRP)-conjugated goat anti-mouse secondary antibody (Jackson ImmunoResearch).

Techniques: Incubation, Infection, Cell Culture, Microscopy, Western Blot, Binding Assay

Journal: PLOS Neglected Tropical Diseases

Article Title: Distinct features of the host-parasite interactions between nonadherent and adherent Trichomonas vaginalis isolates

doi: 10.1371/journal.pntd.0011016

Figure Lengend Snippet: The trophozoites of TH17 isolate were pretreated with DMSO or TPI. A. the trophozoites cultured on a glass slide were examined by IFA using anti-α-tubulin or anti-α-actin antibodies. B. Axostyles in the trophozoites co-cultured with h VECs were observed by SEM. The arrowheads indicate axostyle (Ax); anterior flagellum (AF); recurrent flagellum (RF). C. Protein lysates from treated TH17 trophozoites were subjected to western blotting using anti-α-tubulin (α-Tub) or anti-GAPDH antibodies. D. CFSE-prelabelled trophozoites were co-cultured with h VECs (moi 1:3) for 30 min and the number of trophozoites bound per 1,000 h VECs was calculated as shown in the bar graph (mean ± SD). All assays were repeated three times. Significant differences were statistically analyzed by the Student’s t-tests with P < 0.01(**), P <0.05 (*), and ns, not significant.

Article Snippet: The membrane was sequentially incubated with mouse anti-α-actin (10,000× Ac-40, Genetex), mouse anti-α-tubulin (10,000×, Sigma, DM1A), mouse anti- Tv CyP2 (3,000×) [ ], and mouse anti-GAPDH (5,000×, a gift from Dr. John Alderete, Washington State University) and horseradish peroxidase (HRP)-conjugated goat anti-mouse secondary antibody (Jackson ImmunoResearch).

Techniques: Cell Culture, Western Blot

Journal: International Journal of Biological Sciences

Article Title: Instillation of particulate matter 2.5 induced acute lung injury and attenuated the injury recovery in ACE2 knockout mice

doi: 10.7150/ijbs.23489

Figure Lengend Snippet: Change in ACE and ACE2 expressions in the lungs of mice after PM 2.5 treatment. The pulmonary ACE and ACE2 expression were determined by western blot. β-actin was used as the internal control for ACE and ACE2. The value of ACE and ACE2 expression in non-PM 2.5 treatment was calculated in 100% as the sham value. The relative changes of pulmonary ACE expression in WT and ACE2 KO mice at 2 and 5 days postinstillation (A) . The relative changes of pulmonary ACE2 expression in ACE2 KO mice at 2 and 5 days postinstillation (B) . All values are expressed as the mean ± SD from each group; * p < 0.05 and ** p < 0.01 compared with the Sham value in the same group; † p < 0.05, †† p < 0.01 and ††† p < 0.001 compared with 5 days to 2 days postinstillation in same group; # p < 0.05 compared with the ACE2 KO mice to WT mice in the same day.

Article Snippet: The primary ACE, ACE2, phosphor-STAT3, phosphor-ERK1/2, STAT3, ERK1/2 and β-actin antibodies were obtained from Genetex (Irvine, CA, USA) or Cell Signaling Technology (Beverly, MA, USA).

Techniques: Expressing, Western Blot