Journal: PLoS ONE

Article Title: Identification of Tyrosine-9 of MAVS as Critical Target for Inducible Phosphorylation That Determines Activation

doi: 10.1371/journal.pone.0041687

Figure Lengend Snippet: (A) HEK293T cells were transfected with increasing amount of Flag-vector, Flag-WT or Flag-Y9F mutant. Cells were harvested for Trypan Blue counting 48 h post-transfection. Data are mean of three independent experiments ± SEM done in triplicate. (B) HEK293T cells were transfected with 1.0 µg expression vector encoding Flag-vector, Flag-WT or Flag-Y9F mutant. Annexin V assays were performed 48 hours post-transfection.

Article Snippet: Annexin V staining was performed as indicated with the Annexin V-Biotin Apoptosis Detection Kit (BD Bioscience).

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Expressing

Journal: Molecular Cancer

Article Title: ß3 integrin modulates transforming growth factor beta induced (TGFBI) function and paclitaxel response in ovarian cancer cells

doi: 10.1186/1476-4598-11-36

Figure Lengend Snippet: Suppression of different integrin and ECM components has distinct effects on paclitaxel-induced death in ovarian cancer cells. A, Western blot analysis was performed on SKOV3 cells transfected with either non-target control siRNA, ß1 integrin siRNA, or ß3 integrin siRNA. Cells were subjected to varying concentrations of paclitaxel for 24 hours followed by flow cytometric analysis of Annexin-V and propidium iodide staining, *p < 0.05, **p < 0.01. B, Cell titre glo cell viability assay performed on non-target control, ß1 integrin, or ß3 integrin siRNA expressing cells, treated with increasing concentrations of Paclitaxel for 48 hours. **p < 0.01, ***p < 0.001. C, SKOV3 cells were infected with Lentivirus expressing either non-target control shRNA, TGFBI shRNA, or two separate fibronectin shRNA targets. Western blot analysis was performed on RIPA soluble lysates using antibodies to the specified proteins. Cells were treated with 0.3 μM paclitaxel for 24 hours followed by Annexin-V and propidium iodide staining, and subsequently analyzed by flow cytometry. Experiments were performed in triplicate and are represented as fold increase in cells in early apoptosis, **p < 0.01.

Article Snippet: The TACS Annexin-V Apoptosis kit (R&D systems Europe Ltd.) was performed according to manufacturer’s instructions.

Techniques: Western Blot, Transfection, Control, Staining, Viability Assay, Expressing, Infection, shRNA, Flow Cytometry

Journal: PLoS ONE

Article Title: Hypoxia-activated prodrug TH-302 decreased survival rate of canine lymphoma cells under hypoxic condition

doi: 10.1371/journal.pone.0177305

Figure Lengend Snippet: After treatment with 50 μM TH-302 for 24 h during culture under 5% O 2 , apoptotic cells were evaluated with fluorescent annexin V staining (400× magnification). Light microscope images were obtained after Wright–Giemsa staining (1000× magnification). The percentage of apoptotic cells was quantified with a fluorescent image analyzer, and the data are presented as percentages (%) of the control values. Each bar represents the mean ± SD of three separate experiments. ** P < 0.01 (Dunnett’s test).

Article Snippet: Apoptosis was evaluated with the Annexin V–Biotin Apoptosis Detection Kit (with streptavidin-FITC; Blue Heron Biotechnology, Bothell, WA, USA), according to the manufacturer’s instructions.

Techniques: Staining, Light Microscopy, Control