27 ohc

The 25-OHC, 7-OHC, 4-PBA, and Hoechst 33342 were from Sigma-Aldrich. The 27-OHC and 7-ketocholesterol (7-keto) were from Santa Cruz. The FITC annexin V apoptosis detection kit I was from BD Biosciences. Cell Counting Kit-8 (CCK-8) was from Dojindo.

SH‐SY5Y, a human neuroblastoma cell line, was obtained from the Cell Resource Center, IBMS, CAMS/PUMC, Beijing, China. The C6 rat glioma cell line was obtained from the Type Culture Collection of the Chinese Academy of Sciences, Shanghai, China. The compound, 27‐OHC (540 μg), was purchased from Santa Cruz Biotechnology, USA, and dissolved in 200 μl of dimethylsulfoxide (DMSO) to create a stock solution; the stock solution was then diluted in DMEM to 400 µM and stored at −80°C until use. Cells were cultured in DMEM containing 10% fetal bovine serum and streptomycin (10 U/L) and incubated in 5% CO₂ at 37°C. All experiments were conducted under treatment with various treatments (5, 10, or 20 μM) of 27‐OHC or vehicle (DMEM) for 24 hr, and cells or culture media were then collected for the indicated assay measurements. SH‐SY5H and C6 cells were treated with DMEM or 27‐OHC (5, 10 or 20 μM) for 24 hr and harvested for measurement. The dose range of 27‐OHC was based on our previous study (An et al., 2017; Ma, Li et al., 2015).