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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