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The indicator cells were cultivated overnight with methionine (in the sulfur auxotroph) and GSH (in the gsh1 strain) in the pre-cultures and starved without these sources for 5-6 hours before re-inoculating at an initial OD 600 nm of 0.15 in media containing 100 M of GSH (for the sulfur auxotrophic strain) and no GSH for the glutathione deficient strain

The plates were incubated at 30C for 2 days