human t cell acute lymphoblastic leukemia t all cell line molt4  (ATCC)

Journal: PLoS Neglected Tropical Diseases

Article Title: Mitochondria and lipid raft-located F O F 1 -ATP synthase as major therapeutic targets in the antileishmanial and anticancer activities of ether lipid edelfosine

doi: 10.1371/journal.pntd.0005805

Figure Lengend Snippet: ( A ) L . panamensis promastigotes were untreated (Control) or treated with 10 μM edelfosine at the indicated times, and cells with disrupted ΔΨ m (DiOC 6 (3) low ) and ROS production (HE→Eth) were measured by flow cytometry. The numbers in each quadrant refer to the percentages of cells in each population. ( B ) L . panamensis promastigotes untreated (Control) and treated with edelfosine (EDLF) for 3 h were incubated with 2 μM HE and 10 μg/ml Hoechst 33342, and then analyzed by fluorescence microscopy. ( C ) L . panamensis promastigotes and ( D ) Jurkat cells were preincubated with 10 μg/ml CsA for 1 h, or with 10 mM NAC or 10 mM GSH for 2 h, and then incubated in the absence or presence of 10 μM edelfosine for 9 h. After treatment, the percentage of hypodiploid cells was analyzed by flow cytometry. Untreated control cells were run in parallel. ( E ) L . panamensis promastigotes and ( F ) Jurkat cells were preincubated with 10 μM rotenone, 5 mM malonate, 10 μM antimycin A, 1.5 mM azide, 50 μM CCCP or with 1 and 10 μM oligomycin ( L . panamensis and Jurkat cells, respectively) for 1 h, and then incubated in the absence or presence of 10 μM edelfosine for 9 h. After treatment, cells producing ROS were quantified by flow cytometry. Untreated control cells were run in parallel. Data shown are means ± SD or representative of three independent experiments performed. Asterisks denote that the differences between the indicated groups (C and D) and with control cells (E and F) are statistically significant. (*) P <0.05. (**) P <0.01.

Article Snippet: We also found that edelfosine was very efficient in promoting cell death in additional human leukemic cell lines, including human T-cell acute lymphoblastic leukemia (T-ALL) cell lines Jurkat (53.4 ± 6.2% apoptosis) and CEM-C7H2 (58.2 ± 5.9% apoptosis).

Techniques: Control, Flow Cytometry, Incubation, Fluorescence, Microscopy

Journal: PLoS Neglected Tropical Diseases

Article Title: Mitochondria and lipid raft-located F O F 1 -ATP synthase as major therapeutic targets in the antileishmanial and anticancer activities of ether lipid edelfosine

doi: 10.1371/journal.pntd.0005805

Figure Lengend Snippet: ( A ) L . panamensis promastigotes and T-cell leukemia Jurkat cells were untreated (Control) or pretreated with MCD, and then incubated in the absence or presence of 10 μM edelfosine for 24 h. Percentage of hypodiploid cells were measured by flow cytometry. ( B ) L . panamensis promastigotes and T-cell leukemia Jurkat cells were untreated (Control) or pretreated with MCD and then incubated with 10 μM [ 3 H]edelfosine for 1 h. Drug uptake was determined as shown in the Materials and Methods section. Data shown are means ± SD of three independent experiments performed. Asterisks denote that the differences between the indicated groups are statistically significant. (**) P <0.01. (***) P <0.001.

Article Snippet: We also found that edelfosine was very efficient in promoting cell death in additional human leukemic cell lines, including human T-cell acute lymphoblastic leukemia (T-ALL) cell lines Jurkat (53.4 ± 6.2% apoptosis) and CEM-C7H2 (58.2 ± 5.9% apoptosis).

Techniques: Control, Incubation, Flow Cytometry

Journal: PLoS Neglected Tropical Diseases

Article Title: Mitochondria and lipid raft-located F O F 1 -ATP synthase as major therapeutic targets in the antileishmanial and anticancer activities of ether lipid edelfosine

doi: 10.1371/journal.pntd.0005805

Figure Lengend Snippet: ( A ) Jurkat cells untreated (Control) and treated with 10 μM edelfosine for 9 h were lysed in 1% Triton X-100 and subjected to discontinuous sucrose density gradient centrifugation. Individual fractions were subjected to SDS-PAGE, and location of GM1 was determined using CTx B subunit conjugated with horseradish peroxidase. ( B ) Proteins from lipid rafts of untreated control and edelfosine-treated Jurkat cells were subjected to two-dimensional gel electrophoresis followed by MALDI-TOF analysis. Mitochondrial F O F 1 -ATP synthase β subunit is indicated by an arrow. ( C ) Mass spectrum of the tryptic peptides of the F O F 1 -ATP synthase β subunit spot. Mass value (m/z) and putative amino acid position assignments are indicated above peaks. ( Inset ) Peptide coverage map of human F O F 1 -ATP synthase β subunit; the peptides used for identification are highlighted in bold characters and underlined. ( D ) Jurkat cells were untreated (Control) or preincubated with 10 μM oligomycin for 1 h and then incubated in the absence or presence of 10 μM edelfosine for 9 h, and ΔΨ m disruption (Low ΔΨ m ) and DNA breakdown (hypodiploids) were evaluated. Data shown are means ± SD or representative of three independent experiments. Asterisks denote that the differences between the indicated groups are statistically significant. (**) P <0.01.

Article Snippet: We also found that edelfosine was very efficient in promoting cell death in additional human leukemic cell lines, including human T-cell acute lymphoblastic leukemia (T-ALL) cell lines Jurkat (53.4 ± 6.2% apoptosis) and CEM-C7H2 (58.2 ± 5.9% apoptosis).

Techniques: Control, Gradient Centrifugation, SDS Page, Two-Dimensional Gel Electrophoresis, Electrophoresis, Incubation, Disruption