The level of nitrite in the ARPE19 cells was determined as an indicator of NO production using the Griess reagent as in a previous study [22 (link)]. Briefly, the culture supernatants were collected from subset groups of ARPE19 cells after subsequent treatment with MED, A2E, or BL. These supernatants (100 μL) were mixed with 100 μL of modified Griess reagent (Invitrogen Co., Carlsbad, CA, USA) in 96-well plates. After incubation for 5 min, the absorbance at 540 nm was detected using a VersamaxTM microplate reader (Molecular Devices, San Jose, CA, USA).
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