Curcumin-induced N2a Cell Death Analysis

N2a cells were seeded at a density of 5000 cells/well in 96 well plates. Curcumin-mediated N2a cell death was studied using LIVE/DEAD assay reagents (Invitrogen) as described earlier22 (link). Briefly, cells were serum starved for 2 hours and treated with 10, 25, 50 μM of curcumin or equal volume of DMSO as control for 24 hours. Cells were then incubated in media containing calcein-AM and ethidium homodimer for 30 minutes at room temperature in dark. Random images (3–4) were captured using 40X objective lens in Nikon TS100 inverted microscope supported by NIS elements BR (2.3 version) software with same exposure settings. Number of live (green) and dead (red) cells were counted using ImageJ software.