SMN2 Reporter Assay Protocol

The SMN2 reporter cell line [14 (link), 15 (link)] was cultured in high-glucose DMEM supplemented with 10% (v/v) fetal bovine serum (FBS) and 1X Penicillin/Streptomycin. Reporter cells media was supplemented with hygromycin during maintenance. All cells were maintained at 37°C and 5°C CO₂. After seeding in 96-well white tissue culture plates at 25,000 per well, cells were incubated overnight before being exposed to different doses of compounds or DMSO for 24 hrs. The final DMSO concentration was kept constant at 0.1% (v/v). Firefly and renilla luciferase expression were assayed with DualGlo (Promega E2920) and measured on a PHERAstar FS microplate reader (BMG Labtech). Relative light units are normalized to DMSO control and expressed as a percentage.