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85035169825
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6 PFU), and then immediately given BrdU (Sigma) drinking water (0.8 mg/ml) (4). BrdU water was made fresh and changed daily. LN cells were stained for flow cytometry with mAbs to CD8-PE (Gibco-BRL, Gaithersburg, MD), to CD4-PE (Collaborative Biomedical Products, Bedford, MA), to CD44-biotin (IM7.8.1), or to Ly-6C-biotin (Pharmingen, San Diego, CA); with streptavidin-RED613 (Gibco-BRL); and, after fixation, with mAb to BrdU-FITC (Becton Dickinson, Mountainview, CA) (4). Stained cells were analyzed on a FACScan flow cytometer (Becton Dickinson).
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39
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85035162653
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hi cells in ATx 2C mice, cells were stained with mAb to CD8-CyChrome (Pharmingen), 1B2-biotin (32), Texas Red-streptavidin (Gibco-BRL), and mAb to CD44-PE (Pharmingen) before being stained with mAb to BrdU-FITC. Four-color analysis was done with a FACStar Plus flow cytometer (Becton Dickinson).
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40
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85035165548
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5 U of purified murine IFN-β (Lee Biomolecular Laboratory, San Diego, CA) or an equivalent volume (0.45 ml) of mock IFN α/β (Lee Biomolecular Laboratory) and were immediately started on BrdU drinking water. Staining of LN and spleen cells for flow cytometry was carried out as described in Fig. 1.
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41
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+ cells was calculated from total cell counts of pooled LNs and spleens. Experiments 1 and 2 represent independent experiments.
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42
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85035161077
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note
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We thank M. B. A. Oldstone for advice on viral experiments, D. Y. Loh for 2C mice, I. Gresser for providing antiserum to IFN, T. Knapp for assistance with four-color analysis, and B. Marchand for typing the manuscript. Supported by grants CA38355, CA25803, AI21487, and AI32068 from the USPHS and by a Centennial Fellowship from the Medical Research Council of Canada (to D.F.T.). This is publication 9662-IMM from the Scripps Research Institute.
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