Fig. 1. FreSHtracer-based glutathione regeneration capacity (GRC) assay. (a) A reaction equation for FreSHtracer and glutathione (GSH). F510 (Ex430-510), GSH unbound signal; F580 (Ex520-Em580), GSH bound signal. Fluorescence ratio of FreSHtracer (FR, F510/F580) is proportional to GSH level. (b) Experimental overview for quantifying GRC using FreSHtracer or MitoFreSHtracer. Cells loaded with FreSHtracer or MitoFreSHtracer are observed after treatment with diamide (DA) or N-ethylmaleimide (NEM). (c) GRC was quantified on the basis of the area under the curves (AUC) after non-treatment (A+B+C area), treatment of DA (for GRC, B+C area) and NEM (for the basal thiol level in cells, C area). The ratio of B area and A plus B area is assumed to be GRC.

International Journal of Stem Cells 2023;16:356-62 https://doi.org/10.15283/ijsc23047

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