Journal: Cell Death & Disease

Article Title: Constitutive localization of DR4 in lipid rafts is mandatory for TRAIL-induced apoptosis in B-cell hematologic malignancies

doi: 10.1038/cddis.2013.389

Figure Lengend Snippet: ( a ) Surface expression of TRAIL receptors in transformed and non-transformed cells. Flow cytometry analysis of surface expression level of DR4 and DR5 in four Burkitt lymphoma cell lines. Histograms obtained in a representative experiment are shown. Numbers represent the median fluorescence intensity. Results obtained in a representative experiment are shown. ( b ) Graphs showing the surface expression level of DR4 and DR5 in four Burkitt lymphoma cell lines (left panel) and in non-transformed cells PBL, fibroblasts and HUVEC (right panel). Mean±S.D. of the median fluorescence intensity obtained in four different experiments is reported. ( c ) Soluble TRAIL-induced apoptosis in transformed and non-transformed cells. Biparametric flow cytometry analysis of sTRAIL-induced apoptosis in four Burkitt lymphoma cell lines after double staining with annexin V-FITC/Trypan blue. Numbers represent the percentage of dead cells either annexin V/Trypan blue double-positive or annexin V single-positive. Results obtained in a representative experiment are shown. ( d ) Bar graphs showing the amount of apoptosis in four Burkitt lymphoma cell lines (left panel) and in non-transformed PBL, fibroblasts and HUVEC (right panel) after sTRAIL administration. Mean±S.D. of the percentages of annexin V-positive cells obtained in four different experiments is reported. Significant differences ( P <0.01) were detected between control and treated cancer cells

Article Snippet: Apoptosis was induced by incubating cells with recombinant soluble TRAIL (TRAIL 100 ng/ml for 48 h, Adipogen International, Inc. San Diego, CA, USA) or with agonist antibodies anti-DR4 and anti-DR5 (2 μ g/ml, both R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Transformation Assay, Flow Cytometry, Fluorescence, Double Staining, Control

Journal: Cell Death & Disease

Article Title: Constitutive localization of DR4 in lipid rafts is mandatory for TRAIL-induced apoptosis in B-cell hematologic malignancies

doi: 10.1038/cddis.2013.389

Figure Lengend Snippet: Lipid rafts and TRAIL-induced apoptosis. Left panels. Bar graphs showing the amount of sTRAIL-induced apoptosis in cells pretreated with MBC or perifosine. Mean±S.D. of the percentages of annexin V-positive cells obtained by flow cytometry in four different experiments is reported. Right panels. IVM analysis after triple cell staining with anti-DR4/anti-GM3/Hoechst in control, MBC- or perifosine-treated cells. The yellow fluorescence areas indicate the co-localization. ( a ) Ramos lymphoma cell line, ( b ) Namalwa lymphoma cell line and ( c ) freshly isolated human lymphocytes. * indicates P <0.01 ° indicates P <0.05 versus sTRAIL samples. ( d ) Quantitative evaluation of GM3/DR4 and GM3/DR5 association by FRET technique, as revealed by flow cytometry analysis. Numbers represent the FRET efficiency (calculated by using Riemann algorithm). Note different scales

Article Snippet: Apoptosis was induced by incubating cells with recombinant soluble TRAIL (TRAIL 100 ng/ml for 48 h, Adipogen International, Inc. San Diego, CA, USA) or with agonist antibodies anti-DR4 and anti-DR5 (2 μ g/ml, both R&D Systems, Minneapolis, MN, USA).

Techniques: Flow Cytometry, Staining, Control, Fluorescence, Isolation

Journal: Cell Death & Disease

Article Title: Constitutive localization of DR4 in lipid rafts is mandatory for TRAIL-induced apoptosis in B-cell hematologic malignancies

doi: 10.1038/cddis.2013.389

Figure Lengend Snippet: ( a ) Ex vivo analyses of lymphocytes isolated from patients with CLL. Flow cytometry analysis of surface expression level of CD19 in lymphocytes freshly isolated from a representative HD among six or from two pathological subjects among six (Pt1 and Pt2) affected by B leukemia. Numbers represent the percentage of CD19-positive cells. Results obtained in a representative experiment are shown. ( b ) Surface expression of TRAIL receptors. Flow cytometry analysis of surface expression level of DR4 and DR5 in lymphocytes freshly isolated from a healthy donor or from two pathological subjects affected by B leukemia. Numbers represent the median fluorescence intensity. Results obtained in a representative experiment are shown. ( c ) Apoptosis induction by sTRAIL, mTRAIL and agonist antibodies to DR4 and DR5. Biparametric flow cytometry analysis of apoptosis induced by sTRAIL or by mTRAIL, that is, coculturing CD34 + -armed cells or CD34 + -mock with target cells (1 : 1 ratio). Numbers represent the percentage of apoptotic cells either annexin V/Trypan blue double-positive or annexin V single-positive. Results obtained in a representative experiment are shown. Analyses shown in b and c were restricted to CD19-positive cells

Article Snippet: Apoptosis was induced by incubating cells with recombinant soluble TRAIL (TRAIL 100 ng/ml for 48 h, Adipogen International, Inc. San Diego, CA, USA) or with agonist antibodies anti-DR4 and anti-DR5 (2 μ g/ml, both R&D Systems, Minneapolis, MN, USA).

Techniques: Ex Vivo, Isolation, Flow Cytometry, Expressing, Fluorescence