Journal:

Article Title: Skin-stage Schistosomula of Schistosoma mansoni Produce an Apoptosis-inducing Factor That Can Cause Apoptosis of T Cells *

doi: 10.1074/jbc.M201344200

Figure Lengend Snippet: Following incubation, apoptosis was measured by annexin V and propidium iodide staining and counting 500 –1000 cells under a fluorescence microscope (A), by measuring caspase-3 activity in various subset of lymphocytes using PhiPhiLux substrate and counting 500 –1000 cells under a fluorescence microscope (B), and by evaluating DNA fragmentation in different subsets after treatment with DNA digestion buffer and separation on a 1% agarose gel (C). Data presented are representative of one of three to five similar experiments using five to seven mice per group in each experiment. A and B show means ± S.D. of the percentage of positive cells. * and **, p < 0.05 and p < 0.01 compared with the medium control, respectively.

Article Snippet: Apoptotic cells around the parasites in the skin of mice were evaluated by TUNEL staining using an in situ apoptosis detection kit (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.

Techniques: Incubation, Staining, Fluorescence, Microscopy, Activity Assay, Agarose Gel Electrophoresis, Control

Journal:

Article Title: Skin-stage Schistosomula of Schistosoma mansoni Produce an Apoptosis-inducing Factor That Can Cause Apoptosis of T Cells *

doi: 10.1074/jbc.M201344200

Figure Lengend Snippet: 1 × 106 Thy1.2+ cells isolated from skin-draining lymph nodes of naive mice were cultured with 60 μg/ml ES products or ES products plus 5 μg/ml recombinant Fas-Fc fusion protein for 24 h. Apoptosis was evaluated by determining the percentage of annexin V-positive cells. *, p < 0.01 compared with the medium control.

Article Snippet: Apoptotic cells around the parasites in the skin of mice were evaluated by TUNEL staining using an in situ apoptosis detection kit (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.

Techniques: Isolation, Cell Culture, Recombinant, Control

Journal:

Article Title: Skin-stage Schistosomula of Schistosoma mansoni Produce an Apoptosis-inducing Factor That Can Cause Apoptosis of T Cells *

doi: 10.1074/jbc.M201344200

Figure Lengend Snippet: ES products were divided into three fractions, fraction 1 (Fr 1; <3 kDa), fraction 2 (Fr 2; 3–30 kDa), and fraction 3 (Fr 3; >30 kDa). Bands in fraction 2 were again separated by gel filtration. Each peak (at 60 μg/ml) was then tested for its ability to induce apoptosis of T cells (isolated from the skin-draining lymph nodes of naive mice) by staining for annexin V. Only two peaks (23 and 28 kDa) are shown. About 500 cells were counted under a fluorescence microscope. Values are representative of one of three similar experiments and are means ± S.D. of the percentage of positive cells. * and **, p < 0.05 and p < 0.001 compared with the medium control (M), respectively. ES products from normal schistosomula were used as a positive control.

Article Snippet: Apoptotic cells around the parasites in the skin of mice were evaluated by TUNEL staining using an in situ apoptosis detection kit (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.

Techniques: Filtration, Isolation, Staining, Fluorescence, Microscopy, Control, Positive Control

Journal: Gene therapy

Article Title: Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation.

doi: 10.1038/gt.2008.77

Figure Lengend Snippet: Figure 3 Mouse UL16-binding protein-like transcript 1 E (MULT1E)/FasTI induces apoptosis. A total of 1 106 cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml NKG2D/Fc for 16 h. The cells were then analyzed for apoptosis and necrosis using Annexin V assay (a–c) or caspase-3 assay (d) according to the manufacturers’ protocols. (a) An example of the fluorescence-activated cell sorting (FACS) data. (b, c) Summaries of data from three separate experiments. The statistical analyses were conducted between the controls (open bars) and NKG2D/Fc-treated cells (solid bars) using two-way analysis of variance (ANOVA). The difference between NKG2D/Fc-treated L-5 cells and NKG2D/Fc-treated L-10 cells was also compared using Student’s t-test. *Po0.05; **Po0.01 and ***Po0.001.

Article Snippet: Induction of apoptosis in cells expressing the fusion protein To determine if cells expressing the fusion protein can be induced to undergo apoptosis, one million cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml of NKG2D/Fc for 16 h. Apoptosis of the cells was measured using two systems: a TACS Annexin V-FITC Apoptosis Kit (R&D Systems) and a caspase-3 fluorometric assay (R&D Systems).

Techniques: Binding Assay, Clone Assay, Annexin V Assay, Caspase-3 Assay, FACS

Journal: Journal of biomedical materials research. Part A

Article Title: In vitro evaluation of bone cell response to novel 3D-printable nanocomposite biomaterials for bone reconstruction.

doi: 10.1002/jbm.a.37719

Figure Lengend Snippet: FIGURE 5 Differentiated MC3T3-E1 osteoblast cell proliferation on mSLA-printed nanocomposite discs. SP10 (10% nanohydroxyapatite [nHA]), mSP10 (methacrylated acrylated epoxidized soybean oil [mAESO] and 10% nHA), SP30 (30% nHA), mSP30 (mAESO and 30% nHA), and hydroxyapatite (HA) (control). Proliferation of dMC3T3-OB cells as measured by XTT assay. Data are means ± SD (n = 8 for nanocomposite discs and n = 5 for HA discs, from four experiments), (statistically significant, *p ≤.05, **p ≤.01, ***p ≤.001).

Article Snippet: Cell proliferation after 1, 3, and 7 days of culture was characterized using XTT (sodium 30-[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis (4-methoxy6-nitro) benzene sulfonic acid hydrate; TACS XTT cell proliferation kit, R&D Systems, Minneapolis, USA).

Techniques: Control, XTT Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Induction of alloantigen-specific human T regulatory cells by vasoactive intestinal peptide.

doi: 10.4049/jimmunol.0900400

Figure Lengend Snippet: FIGURE 1. VIP induces cell cycle arrest and inhibits cytokine-producing potential of allogeneic stimulated PBMCs. A, VIP was administered at different doses to a primary MLC with allogeneic human PBMCs. Prolifer- ation was determined after 5 days of culture. Results are expressed as percentage of inhibition of proliferation relative to untreated cultures. B, Primary MLCs with allogeneic human PBMCs were treated with medium (control) or VIP (107 M). Proliferation was determined at different time points after initiation of culture. C, Ap- optotic cells were determined at 48 h by flow cytometry after propidium iodide and annexin V staining. Cells incubated with medium alone were used as unstimu- lated naive cells. Numbers represent percentage of cells in each quadrant. D, Percentages of cells in G0/G1 and S phases (on gated CD4 T cells) were determined at different time points after initiation of MLC. E, The cy- tokine-producing potential was determined by intracel- lular cytokine staining using flow cytometry analysis on gated CD4 T cells after 48 h of MLC. Numbers rep- resent the percentage of cytokine-positive cells. Fluo- rescence intensity is plotted against the forward scatter of the cells (FSC). n 4–5 experiments performed in duplicate. , p 0.001 vs control.

Article Snippet: Assessment of apoptosis and cell viability Quantitative determination of viability was performed using an annexin V-based apoptosis detection kit (R&D Systems) and subsequently analyzed by flow cytometry.

Techniques: Inhibition, Control, Cytometry, Staining, Incubation