Journal:

Article Title: Adenosine A 2A receptor activation protects CD4 + T lymphocytes against activation-induced cell death

doi: 10.1096/fj.10-155192

Figure Lengend Snippet: A, B) AICD of T cells is mediated by Fas-FasL interactions. Murine 7.5 T-hybridoma cells were stimulated with ConA in the presence of 10 μg/ml FLIM58 anti-mouse FasL (a-FasL) neutralizing antibody or its isotype control. After 18–20 h, cell viability was assessed using the MTT (A) and LDH (B) assays. Results are means ± se of a representative experiment from 3 separate experiments; n = 5 wells/group. ** P < 0.01. C–E) Selective A2AR agonist CGS21680 (CGS) decreases FasL mRNA levels in ConA-stimulated murine 7.5 hybridoma (C) and human Jurkat (D) T-cell lines, and also decreases Fas mRNA levels in ConA-stimulated 7.5 murine T hybridoma cells (E). Cells were stimulated with ConA for 6 h, and CGS (100 nM) or its vehicle (veh) was added to the cells 30 min before stimulation. Fas and FasL mRNA levels were determined using real-time PCR. Results are means ± se of one representative experiment from 3 separate experiments; n = 4 samples/group. **P < 0.01; ***P < 0.001. F) CGS decreases the activation-induced expression of Fas protein levels in T cells. Murine 7.5 T-hybridoma cells were treated with 100 nM CGS or vehicle (V) 30 min before activation by ConA. Whole cell proteins were extracted at various time points after stimulation and analyzed by Western blotting. β-Actin, used as an internal control, is shown in Fig. 4. Western blots in one figure were done on the same membrane. One representative experiment of 3 is shown. unstim, unstimulated; M, medium.

Article Snippet: FLIM58 anti-mouse FasL neutralizing antibody was purchased from MBL International (Woburn, MA, USA), and hamster IgG isotype control was from BD Pharmingen (San Diego, CA, USA).

Techniques: Control, Real-time Polymerase Chain Reaction, Activation Assay, Expressing, Western Blot, Membrane

Journal:

Article Title: Matrix metalloproteinase-7 is expressed by pancreatic cancer precursors and regulates acinar-to-ductal metaplasia in exocrine pancreas

doi: 10.1172/JCI15051

Figure Lengend Snippet: Expression of Fas and FasL after duct ligation. Immunoreactivity for Fas (a) and FasL (b) in wild-type pancreata 5 days after ductal ligation. Bar represents 50 μm. Arrows in a indicate positive staining of metaplastic ducts among large regions of positively staining acinar cells. (c) Immunoblot of FasL in lysates isolated at 0, 5, 7, and 10 days after ductal ligation, using a rabbit polyclonal anti-FasL. Numbers indicate size markers. Full-length FasL and sFasL are indicated. (d) Immunoblot for FasL in pancreatic lysates isolated from wild-type and MMP7–/– mice 10 days after ductal ligation, using a rat monoclonal anti-mouse FasL. Protein loading in c and d was confirmed by Fast Green stain of the blot (Load).

Article Snippet: Antibodies and dilutions were as follows: rabbit polyclonal anti–MMP-7 antibody ( 11 ) was used at 1:200; rabbit polyclonal anti-FasL (SC-6327; Santa Cruz Biotechnology Inc., Santa Cruz, California, USA) was used at 1:500; and rat monoclonal anti-mouse FasL antibody (Alexis Biochemicals, San Diego, California, USA) was used at 1:500.

Techniques: Expressing, Ligation, Staining, Western Blot, Isolation