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After the treatment of cells with the tested compounds, 20 L of MCB solution was added to the cells in 96-well plates and the measurement started immediately (final concentration of MCB in a well was 40 M)

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The reaction between RT and GSH is based on a Michael addition reaction that is inherently reversible, has an appropriate dissociation equilibrium constant K d in the mM range and rapid reaction kinetics, and provides ratiometric readouts, thus allowing GSH quantification independent of the probe concentration