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10544249382
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note
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6 melanoma cells were separated by SDS-PAGE (10%) under nonreducing conditions and immunoblotted (ECL system, Amersham) using the affinity-purified rabbit anti-FasL antibody PE62 (33).
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note
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5 labeled cells in a total volume of 200 μ1.
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10544230136
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2 and 0.1% azide. Supernatants (1:1 dilution) of the monoclonal antibody (mAb) to FasL, A11 (33) (Alexis Corp., San Diego, CA) was added to the section for 1 hour, and after rinsing in PBS, peroxidase-conjugated goat antibody to rat Ig (TAGO, Burlingame, CA) was added for 0.5 hour. After rinsing, antibody location was revealed by the AEC chromogen. For the detection of melanoma cells, a mouse mAb to MAGE-1, detecting a subpopulation of melanoma cells (40), was used (20 μg/ml), and detected with a biotinylated goat anti-mouse Ig and avidin-peroxidase-conjugate (TAGO, Burlingame, CA). Fas antigen was detected with a mouse mAb to human Fas (10 μg/ml). In the negative control, the FasL antibody was replaced with rat Ig. The mAb to human melanoma (HMB45) was from ENZO Diagnostics Inc., New York, USA.
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0030030950
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10544250993
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note
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Frozen sections were stained as in Fig. 2. T cells were detected using the anti-CD3 specific antibody leu-4 (Becton-Dickinson, Oxnard, CA). TUNEL staining was done according to kit instructions from Boehringer, Mannheim, Germany.
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10544252202
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note
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5, American Type Tissue Culture) were injected subcutaneously into 4-week old C57BL/6, CSTBL/6-lpr, C57Bl/6-gld (Jackson Laboratory, Bar Harbor, ME), MRL and MRL-lpr mice (Harian, Zeist, Nederland).
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40
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0029812081
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note
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We thank C. Kamel, F. Beerman, and K. Burns for helpful discussions; S. Hertig and S. Aslan for technical and secretarial assistance; and P. H. Krammer and F. Lejeune for reagents. Supported by grants of the Swiss National Science Foundation (J.T.) and of Human Frontier Science (M.H.). This study is dedicated to Dr. Stephan Carrel (Ludwig Institut of Cancer Research, Lausanne branch). With his death, melanoma research has lost a highly noted authority.
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