Oligomeric aβ1 42

Oligomeric Aβ₁₋₄₂ (Sigma Aldrich, USA) was prepared according to manufacturer specifications. Briefly, Aβ₁₋₄₂ was dissolved in 0.2% NH₄OH at 1 mg/mL, brought to 400 nM concentration in ACSF and sonicated for 1 min. Prior to experiments, solution was allowed to fibrillize for minimum of 1 h.

ARPE-19 cells were obtained from the American Type Tissue Culture Collection and cultured in Dulbecco’s modified Eagle’s medium containing the F-12 nutrient mixture (DMEM/F12) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 µg/mL streptomycin. Cultures were maintained at 37 ℃ in an atmosphere of 5% CO₂ and 95% air. All cell culture reagents were purchased from Gibco (Grand Island, MA, USA).
Cultures in the logarithmic phase were used in the experiments described below. Depending on the experiment, cells were treated with the following reagents (Sigma-Aldrich, St. Louis, MO, USA): sulforaphane (1–50 μM), oligomeric Aβ_1-42 (0.1–10 μM), the SIRT1 agonist SRT1720 (0.1, 1, and 5 μM; Sigma-Aldrich), or the SIRT1 inhibitor nicotinamide (2 μM; Sigma-Aldrich). The comprehensive flow sheet abstract was shown in Graphical Abstract.

Sulforaphane, oligomeric Aβ_1-42, the SIRT1 agonist SRT1720, and the SIRT1 inhibitor nicotinamide were purchased from Sigma-Aldrich (St. Louis, MO, USA). Lipofectamine 2000 was provided by Sigma-Aldrich. The antibodies against cleaved caspase-3 (catalog no. #ab32043), cleaved caspase-3 (ab2324), PARP1 (ab32064), SIRT1 (ab189494), and β-actin (ab7817) were obtained from Abcam (UK).