Fclariostar plate reader

After treatment for 24 h, cells were collected on iced PBS. GSH content in MJD26 and MJD78 cells was measured using a GSH + GSSG/GSH assay kit (abcam ab239709; Cambridge, MA, USA) according to the manufacturer’s instructions. Colorimetric intensity at 405 nm was measured using a FCLARIOstar Plate Reader (BMG LabTech, Ortenberg, Germany).

After treatment for 24 h, cells were collected on iced PBS. Catalase activity was assayed in MJD26 and MJD78 cells using AmpliteTM Fluorimetric Catalase assay kits (AAT Bioquest 11306, Sunnyvale, CA, USA) according to the manufacturer’s instructions. Fluorescence intensity (Ex/Em 540/590 nm) was measured using a FCLARIOstar Plate Reader (BMG LabTech, Ortenberg, Germany).

Levels of mitochondrial ROS, total ROS, and protein aggregation in MJD26 and MJD78 cells were determined using MitoSOX Red reagent (M36008, Invitrogen), CellROX™ Orange Flow Cytometry Assay Kits (C10493, Invitrogen), and PROTEOSTAT^® Protein aggregation assays, respectively. After treatment for 24 h, cells were washed with PBS, trypsinized with 0.05% trypsin, and collected into PBS. After treating 1 × 10⁵ cells/mL with 5 μM MitoSOX Red and 500 nM CellROX™ Orange reagent, cells were incubated for 10 min at 37 °C while protected from light. Cells were washed gently 3× with PBS. Fluorescence intensity was measured using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) and an FCLARIOstar Plate Reader (BMG LabTech, Ortenberg, Germany).