Development of a Golgi apparatus-targeting probe for revealing the fluctuations of GSH in the living cells during ferroptosis

Ferroptosis is a distinctive iron-dependent and GSH-related cell death form, and has been demonstrated to closely relate with Golgi apparatus (GA). Exploring the fluctuations of GSH level in GA during ferroptosis is of great importance for the deep research of biological functions of GA in ferroptosis. Herein, we reported a novel GAtargeting probe (NA-G) for the detection of GSH in GA during ferroptosis. The probe NA-G utilized Noxidized pyridine sulfonyl as a responsive site for GSH, with tetradecanoyl amide serving as GA-targeting moiety. NA-G exhibited remarkable sensitivity and selectivity, manifesting a heightened response to GSH and effectively visualizing GSH through bright green fluorescence under UV light. By fluorescence imaging of cellular GSH, NAG was demonstrated to serve as a GA-targeting probe, and could successfully monitor the dynamic changes in intracellular GSH. Further experimentation revealed that NEM stimulation resulted in the removal of a significant amount of endogenous GSH in GA, and erastin-induced ferroptosis significantly reduced GSH levels in GA. Notably, we discovered that Fer-1 and VE effectively inhibited the substantial depletion of GSH in GA during ferroptosis.