HEK293T cells were transfected with an IFN-β luciferase reporter, pRL-CMV (10 ng; Promega) Renilla luciferase control reporter, pcDNA 3.1 and indicated vectors and 24 h later dual luciferase assays (DLAs) were performed as per published protocols60 (link),61 (link). For CpG response assays, TLR9-HEK239s (Invivogen) were transfected using lipofectamine with indicated vectors, IFNβ-luciferase and renilla reporters before being stimulated after 24 h with 5 µg/ml CpG (ODN 2006, InvivoGen). The Burkitt’s lymphoma cell line Ramos was nucleofected with the IFN-β luciferase reporter and pRL-CMV using the NEON transfection system (ThermoFisher Scientific) according to the manufacturer’s instructions.
Jiang S.H., Athanasopoulos V., Ellyard J.I., Chuah A., Cappello J., Cook A., Prabhu S.B., Cardenas J., Gu J., Stanley M., Roco J.A., Papa I., Yabas M., Walters G.D., Burgio G., McKeon K., Byers J.M., Burrin C., Enders A., Miosge L.A., Canete P.F., Jelusic M., Tasic V., Lungu A.C., Alexander S.I., Kitching A.R., Fulcher D.A., Shen N., Arsov T., Gatenby P.A., Babon J.J., Mallon D.F., de Lucas Collantes C., Stone E.A., Wu P., Field M.A., Andrews T.D., Cho E., Pascual V., Cook M.C, & Vinuesa C.G. (2019). Functional rare and low frequency variants in BLK and BANK1 contribute to human lupus. Nature Communications, 10, 2201.
Corresponding Organization : Shanghai Jiao Tong University
Other organizations :
Translational Health Science and Technology Institute, Trakya University, University of Zagreb, University Children's Hospital, Belgrade, Institutul Clinic Fundeni, Children's Hospital at Westmead, Monash University, Walter and Eliza Hall Institute of Medical Research, Fiona Stanley Hospital, Hospital Infantil Universitario Niño Jesús, National Computational Infrastructure
Transfection of HEK293T cells with an IFN-β luciferase reporter, pRL-CMV (10 ng; Promega) Renilla luciferase control reporter, pcDNA 3.1 and indicated vectors
Transfection of TLR9-HEK239s (Invivogen) with indicated vectors, IFNβ-luciferase and renilla reporters
Nucleofection of the Burkitt's lymphoma cell line Ramos with the IFN-β luciferase reporter and pRL-CMV using the NEON transfection system (ThermoFisher Scientific)
dependent variables
Dual luciferase assays (DLAs) performed 24 hours after transfection of HEK293T cells
CpG (ODN 2006, InvivoGen) stimulation of TLR9-HEK239s after 24 hours of transfection
control variables
PRL-CMV (10 ng; Promega) Renilla luciferase control reporter
positive controls
None specified
negative controls
None specified
Annotations
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