HeLa cells (5000 cells/well in 96 well format) were exposed to l-Buthionine-sulfoximine (BSO). BSO inhibits GSH synthesis thus reducing cellular GSH level.
20hr BSO: Cells treated with BSO and assayed for GSH after 20 hours.
BSO + Recovery: Cells treated with BSO for 20 hours then washed 2X with PBS and covered with fresh media without BSO. Cells assayed for GSH after 40 hours.
40hr BSO: Cells treated with BSO, washed at 20 hours and fresh media plus BSO added. Cells assayed for GSH after 40 hours.