ARPE−19 cells were purchased from the American Type Culture Collection (Manassas, VA, USA), and cultured in T25 flasks using 10%FBS, 1% penicillin-streptomycin(p/S) solution in DMEM/F12 basic culture medium (Gibco, Grand Island, NY, USA) at 37 °C and 5% CO2.
In all experiments, ARPE−19 cells used were 4–10 passages. Cells were seeded in 6-well (2 × 105 cells/well), 12-well (1 × 105 cells/well), or 96-well (5 × 103 cells/well) plate. After 12 h, the confluence reached 80–90%, and cells were washed with DPBS (Gibco, Grand Island, NY, USA). Cells were starved in Fetal Bovine Serum (FBS)-free DMEM/F12 (without phenol red) culture medium (Gibco, Grand Island, NY, USA) for 6 h. CAPE or t-BHP was also diluted in FBS-free DMEM/F12 (without phenol red) culture medium. ARPE−19 cells were pretreated with 200 μM Genepin for 2 h. After 2 h 20 μM CAPE treatment, ARPE−19 cells were stimulated with 200 μM t-BHP accordingly. Other reagents (CGA, TCA, IQT, and CUM) were diluted and applied in the same way as CAPE.
In all experiments, ARPE−19 cells used were 4–10 passages. Cells were seeded in 6-well (2 × 105 cells/well), 12-well (1 × 105 cells/well), or 96-well (5 × 103 cells/well) plate. After 12 h, the confluence reached 80–90%, and cells were washed with DPBS (Gibco, Grand Island, NY, USA). Cells were starved in Fetal Bovine Serum (FBS)-free DMEM/F12 (without phenol red) culture medium (Gibco, Grand Island, NY, USA) for 6 h. CAPE or t-BHP was also diluted in FBS-free DMEM/F12 (without phenol red) culture medium. ARPE−19 cells were pretreated with 200 μM Genepin for 2 h. After 2 h 20 μM CAPE treatment, ARPE−19 cells were stimulated with 200 μM t-BHP accordingly. Other reagents (CGA, TCA, IQT, and CUM) were diluted and applied in the same way as CAPE.