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Volumn 280, Issue 5365, 1998, Pages 905-908

Thymocyte development in the absence of pre-T cell receptor extracellular immunoglobulin domains

Author keywords

[No Author keywords available]

Indexed keywords

PROTEIN PRECURSOR; T LYMPHOCYTE RECEPTOR;

EID: 0032496362     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.280.5365.905     Document Type: Article
Times cited : (163)

References (48)
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    • note
    • T were cloned into the p1017 expression vector, which includes the Lck proximal promoter and the human growth hormone 3′-untranslated region (42).
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    • note
    • -/- blastocysts to produce chimeric mice according to standard procedures (43).
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    • 6) were electroporated in 300 μl of serum-free RPMI 1640 medium containing 5 μg of NFAT-luciferase plasmid (44), 6 μg of v-Ras plasmid, and a total of 25 ng of pEF-Boss-based plasmids in a Bio-Rad Gene Pulser set at 240 V and 960 μF. About 6 hours after transfection, the cells were counted in a hemocytometer and plated in triplicate into 96-well plates at 200,000 cells per well. At selected times, the 96-well plates were frozen at -70°C for later analysis of luciferase activity as previously described (27).
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    • note
    • 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.)
    • -/- 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.).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.