NFAT Reporter—HEK293 cell line and Luciferase assay

VP Vasiliki Panagiotakopoulou
DI Dina Ivanyuk
SC Silvia De Cicco
WH Wadood Haq
AA Aleksandra Arsić
CY Cong Yu
DM Daria Messelodi
MO Marvin Oldrati
DS David C. Schöndorf
MP Maria-Jose Perez
RC Ruggiero Pio Cassatella
MJ Meike Jakobi
NS Nicole Schneiderhan-Marra
TG Thomas Gasser
IN Ivana Nikić-Spiegel
MD Michela Deleidi
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The NFAT Reporter—Hek293 cell line was purchased from BPS Bioscience and cultured in DMEM (Merck) supplemented with 10% (vol/vol) FBS (Gibco) and 400 μg/mL of G418. Where specified, cells were transfected with plasmid DNA (SF-tagged wt LRRK2 or SF-tagged LRRK2 G2019S) using 1 mg/mL polyethylenimine (PEI) solution (2.5 μg of plasmid DNA/μL; Polysciences). Where specified, cells were treated with 10 μM colchicine for 30 min at 37 °C or 100 nM PTX for 24 h. Cells were stimulated with 40 ng/mL PMA (Sigma-Aldrich) and 1 μM ionomycin (Sigma-Aldrich) for 24 h. Luciferase activity was determined using the Luciferase Assay System (Promega) according to the manufacturer’s protocol and measured with a TriStar2 S LB 942 Multimode Microplate Reader (Berthold).

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