Journal: Journal of Virology

Article Title: Novel Human Polyomavirus Noncoding Control Regions Differ in Bidirectional Gene Expression according to Host Cell, Large T-Antigen Expression, and Clinically Occurring Rearrangements

doi: 10.1128/JVI.02231-17

Figure Lengend Snippet: Cell lines transfected with NCCR reporter constructs

Article Snippet: Red and green fluorescence levels were measured by flow cytometry and normalized using BKPyV-NCCR(ww) as the reference, with BKPyV EVGR as 1 and BKPyV LVGR as 100 (SD) as detailed in Materials and Methods. table ft1 table-wrap mode="anchored" t5 TABLE 3 caption a7 Cell line Culture medium (supplemented with 10% FBS, 2 mM glutamine) Transfection reagents Origin Cell line source HEK293 DMEM-H Lipofectamine 2000 Human embryonic kidney cells ATCC CRL-1573 HEK293T DMEM-H Lipofectamine 2000 Human embryonic kidney cells expressing SV40 LTag and sTag ATCC CRL-3216 HEK293TT DMEM-H plus 400 μg/ml hygromycin B Lipofectamine 2000 Human embryonic kidney cells expressing SV40 LTag and sTag NCI-Frederick Repository Service, Frederick, MD, USA HEK293MCT DMEM-H Lipofectamine 2000 Human embryonic kidney cells expressing MCPyV LTag and sTag Gift from Nicole Fischer from Institute for Microbiology and Virology, University Medical Center Eppendorf, Hamburg, Germany A375 DMEM-H Lipofectamine 2000 Human malignant melanoma cells ATCC CRL-1619 A549 EMEM Lipofectamine 3000 Human epithelial lung adenocarcinoma ATCC CCL-185 HeLa NT DMEM-H Lipofectamine 2000 Human cervical carcinoma cells Gift from Natalia Teterina from Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA HS683 DMEM-H Omni avalanche Human brain glioma cells ATCC HTB-138 SW480 EMEM Lipofectamine 2000 Human colon adenocarcinoma cells ATCC CCL-228 Open in a separate window Cell lines transfected with NCCR reporter constructs To investigate whether or not expression of SV40 LTag in trans would be able to further increase the host cell-specific expression levels, A375 skin and SW480 colon-derived cells were transfected with a corresponding LTag expression vector, pRcCMV-SLT or pSG-largeT (data not shown).

Techniques: Transfection, Expressing