Retinoic acid (R2625; Sigma) powder was dissolved in DMSO at 3 mg/mL (0.01 M) as stock solution and stored at −80°C with light-protected vials. To induce differentiation of SH-SY5Y cells, we diluted the stock solution with a tissue culture medium at a final concentration of 10 µM.
For the construction of damage models in cellular experiments, we treated SH-SY5Y cells with H2O2 (50 µM). Cells were harvested and followed by subsequent experiments after being treated with H2O2 for 8 hours.
For cells, Amlexanox and Degrasyn were diluted in DMSO according to the manufacturer's instructions and then further diluted in complete cell culture media at a final concentration of 100 µM (Amlexanox) or 5 µM (Degrasyn). For animals, mice are randomly assigned to 2 groups: one group received Degrasyn (40 mg/kg, intraperitoneally) in 100 µL DMSO every 2 days (7 injections); and the other received DMSO (vehicle) every other day (7 injections). CHX (100 µg/mL) was used for protein stability assay and cells were harvested at different time points for the preparation of WCL and subsequent Western blot analysis.
For the construction of damage models in cellular experiments, we treated SH-SY5Y cells with H2O2 (50 µM). Cells were harvested and followed by subsequent experiments after being treated with H2O2 for 8 hours.
For cells, Amlexanox and Degrasyn were diluted in DMSO according to the manufacturer's instructions and then further diluted in complete cell culture media at a final concentration of 100 µM (Amlexanox) or 5 µM (Degrasyn). For animals, mice are randomly assigned to 2 groups: one group received Degrasyn (40 mg/kg, intraperitoneally) in 100 µL DMSO every 2 days (7 injections); and the other received DMSO (vehicle) every other day (7 injections). CHX (100 µg/mL) was used for protein stability assay and cells were harvested at different time points for the preparation of WCL and subsequent Western blot analysis.