Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Effects of interferon gamma on native human acute myelogenous leukaemia cells

doi: 10.1007/s00262-006-0159-1

Figure Lengend Snippet: Effect of IFNγ on in vitro proliferation of AML cells incubated in transwell culture together with either microvascular endothelial cells (EC), HFL1 fibroblasts (FB), Cal72 osteoblastic sarcoma cells (OB) or normal bone marrow stromal cells (BMSC). The figure compares leukaemia cell proliferation for cultures with (+) and without (−) IFNγ 50 ng/ml. Only those samples showing detectable proliferation (i.e. >1,000 cpm) for at least one of the in vitro models (+/−) are presented

Article Snippet: Human lung microvascular endothelial cells were obtained as frozen vials (Cambrex Bio Science Walkersville, Walkersville, MD, USA) and stored in liquid nitrogen until used.

Techniques: In Vitro, Incubation

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Effects of interferon gamma on native human acute myelogenous leukaemia cells

doi: 10.1007/s00262-006-0159-1

Figure Lengend Snippet: The effect of IFNγ on in vitro proliferation of nonleukaemic cells cocultured with AML blasts in transwell cultures. Microvascular endothelial cells (EC), HFL1 fibroblasts (FB) or Cal72 osteoblastic sarcoma cells (OB) were cultured together with AML blasts in the presence (+) or absence (−) of IFNγ 50 ng/ml. Only those patient samples showing detectable proliferation (i.e. >1,000 cpm) for at least one of the in vitro culture models (+/−) are presented

Article Snippet: Human lung microvascular endothelial cells were obtained as frozen vials (Cambrex Bio Science Walkersville, Walkersville, MD, USA) and stored in liquid nitrogen until used.

Techniques: In Vitro, Cell Culture

Journal: The European respiratory journal

Article Title: FTY720-Induced Human Pulmonary Endothelial Barrier Enhancement is Mediated by c-Abl

doi: 10.1183/09031936.00047810

Figure Lengend Snippet: A) HPAEC were stimulated with FTY720 (1 µM) or vehicle control for 15, 30, or 60 min. After treatment, c-Abl was immunoprecipitated with anti-c-Abl (8E9) antibody and incubated with Crk-GST in kinase buffer as described in Methods. The tyrosine phosphorylation of Crk-GST was detected by Western blot as a measure of c-Abl activity. The bar graph represents pooled densitometric results relative to vehicle control from 3 independent experiments. *p<0.01 vs. vehicle control. B) HPAEC or HLMVEC transfected with c-Abl siRNA or control siRNA (si-con) were plated on gold microelectrodes and then stimulated with FTY720 (1 µM). Bar graphs represent the maximal TER obtained after FTY720 relative to baseline resistance (mean ± S.E., n = 3 per condition). *, P<0.05 compared to control siRNA. Western blot demonstrating representative downregulation of c-Abl by siRNA is shown in Suppl. Fig. 1. C) HPAEC were transfected with 100 nM c-Abl or control siRNA and seeded onto Transwell inserts. After FTY720 stimulation (1 µM), FITC-dextran was added into the top chamber and incubated for 2 h. The fluorescent intensity of the bottom chamber was analyzed by fluorometry as per Methods. n = 4. *p<0.031 vs. control siRNA. D) HPAEC were preincubated for 1 h with the c-Abl inhibitor, AG957 (20 µM) or vehicle control and then stimulated with FTY720 (1 µM) or vehicle. Data are a representative of 3 independent experiments.

Article Snippet: Human pulmonary artery endothelial cells (HPAEC) and pulmonary microvascular cells (HLMVEC) (Lonza, Walkersville, MD) were cultured in EBM-2 complete medium (Lonza) with 10% FBS at 37°C in a humidified incubator with 5% CO 2 as previously described [ 4 ].

Techniques: Control, Immunoprecipitation, Incubation, Phospho-proteomics, Western Blot, Activity Assay, Transfection