Journal: Journal of Inflammation (London, England)

Article Title: Oxidative stress increases Fas ligand expression in endothelial cells

doi: 10.1186/1476-9255-3-11

Figure Lengend Snippet: Membrane FasL expression on HUVECs . HUVECs were incubated for 16 hours with or without 1–100 μM H 2 O 2 or 0.5% and 0.5 and 1% CSE, in the presence or absence of 500 U/ml catalase. Membrane FasL expression on HUVECs analyzed by cell-monolayer-based spectrofluorimetry ( A and B ) and flow cytometry ( C ). ( A and B ) Data are reported as the means ± SEM of the values obtained from 5 experiments. *p < 0.05, **p < 0.01 vs. cells not exposed to H 2 O 2 or CSE. †p < 0.05 vs cells exposed to 1% CSE only. ( C ) Data are representative of 3 separate experiments. Background ; fluorescence levels generated by replacement of anti-FasL antibody with nonimmune IgG.

Article Snippet: After blocking with 3% bovine serum albumin and 2% goat serum, sections were incubated with polyclonal rabbit anti-FasL antibody (Wako Pure Chemical Industries, Ltd., Osaka, Japan) or non-immune rabbit IgG (Dako Cytomation) for 1 hour at room temperature.

Techniques: Expressing, Incubation, Flow Cytometry, Fluorescence, Generated

Journal: Journal of Inflammation (London, England)

Article Title: Oxidative stress increases Fas ligand expression in endothelial cells

doi: 10.1186/1476-9255-3-11

Figure Lengend Snippet: Immunohistochemistry of thoracic aortas . Thoracic aortas were excised from rats and exposed to ( B , C ) or not exposed to ( A ) 100 μM H 2 O 2 in vitro for 16 hours. Tissue sections were immunostained with rabbit polyclonal anti-FasL antibody, and immunoreactants were visualized with a substrate solution of 3-amino-9-ethylcarbazole. Cell nuclei were counterstained with a hemtoxylin solution. Exposure of thoracic aortas to 100 μM H 2 O 2 increased expression of FasL on the aortic endothelium ( B ). Arrows point to positive FasL staining (red color in the original photomicrograph). Replacement of anti-FasL antibody with non-immune rabbit IgG resulted in negative staining ( C ). Photographs are representative of immunostaining of three thoracic aorta specimens Original magnification, 200×.

Article Snippet: After blocking with 3% bovine serum albumin and 2% goat serum, sections were incubated with polyclonal rabbit anti-FasL antibody (Wako Pure Chemical Industries, Ltd., Osaka, Japan) or non-immune rabbit IgG (Dako Cytomation) for 1 hour at room temperature.

Techniques: Immunohistochemistry, In Vitro, Expressing, Staining, Negative Staining, Immunostaining

Journal: Journal of Biomedical Science

Article Title: A recombinant scFv-FasL ext as a targeting cytotoxic agent against human Jurkat-Ras cancer

doi: 10.1186/1423-0127-20-16

Figure Lengend Snippet: Immunochemical characterization of cc49scFv-FasL ext and L6scFv-FasL ext . ( A ) Cytotoxicity was detected against A20 target by using culture supernatants from cc49scFv-FasL ext and L6scFv-FasL ext transfected BW5147 and 3T3, respectively. Culture supernatants from BW5147 and 3T3 had no detectable cytotoxicity against A20. ( B and C ) Membrane filtration analysis of cytotoxic materials: A 300 kDa membrane molecular filtration was used as described in Methods. Flow-through and “Retentate” of cc49scFv-FasL ext ( B ) and L6scFv-FasL ext ( C ) samples were readjusted to original volume and tested for cytotoxicity against A20. ( D ) Polyclonal rabbit anti-FasL (C-20) and rabbit anti-HA were used to probe for the presence of fusion proteins. Dilutions used were 1000 fold for anti-HA, 1000 fold for anti-C-20, and 4000 fold for horseradish peroxidase-conjugated anti-rabbit Ab.

Article Snippet: Membranes were then blotted with rabbit polyclonal anti-HA or rabbit polyclonal anti-FasL (C-20) (Santa Cruz Biotechnology) followed by HRP-conjugated donkey anti-rabbit Ab [HRP] (Amersham Biosciences).

Techniques: Transfection, Filtration

Journal: Scientific reports

Article Title: Candidate protein biomarkers in pancreatic neuroendocrine neoplasms grade 3.

doi: 10.1038/s41598-020-67670-7

Figure Lengend Snippet: Figure 3. PanNEN G3 immunostained for FASLG. (a), (b) and (c) FASLG immunoreactivity in membrane of tumour cells. (d) FASLG immunoreactive immune cells infiltrating a non-immunoreactive tumour. Scale bar 100 µm.

Article Snippet: The primary rabbit polyclonal antibody against FAS ligand (FASLG) (HPA054959, Atlas Antibodies, Stockholm, Sweden) was diluted in 1:800 UltraAb Diluent (Thermo Fisher Scientific) followed by incubation for 30 min at room temperature (RT).

Techniques: Membrane