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Single-cell suspensions of thymocytes were stained with the following antibodies (purchased from Caltag, Pharmingen, Becton-Dickinson, or VWR) in phosphate-buffered saline plus 0.3% bovine serum albumin: antibody to CD4 (anti-CD4) (CT-CD4-Tricolor), anti-CD8 [53-6.7-fluorescein isothiocyanate (FITC)], anti-TCRβ [H57-597-phycoerythrin (PE)], anti-CD25 (7D4-FITC), anti-Flag (M2), anti-Myc epitope (9E10), and anti-CD3ε (145-2C11-PE). Jurkat transfectants were stained with Leu 4 (anti-human CD3ε), followed by FITC-conjugated goat antibody to mouse IgG. Stained cells were analyzed with a Becton-Dickinson FACScan and CellQuest software. Dead cells were excluded by gating on the forward and side scatter.
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-/- ES cells; and J. D. Scarborough for help with the production of transgenic mice. Supported by the Irvington Institute (B.A.I.), the Lucille P. Markey Foundation through the University of California, San Francisco, Program in Biological Sciences (N.K.), and the Howard Hughes Medical Institute (F.W.A.)
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-/- ES cells; and J. D. Scarborough for help with the production of transgenic mice. Supported by the Irvington Institute (B.A.I.), the Lucille P. Markey Foundation through the University of California, San Francisco, Program in Biological Sciences (N.K.), and the Howard Hughes Medical Institute (F.W.A.).
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