Wang, Yi-ChuanYi-ChuanWangWu, Ping-HsunPing-HsunWuTing, Wen-ChiehWen-ChiehTingWang, Yi-FuYi-FuWangYang, Ming-HanMing-HanYangSu, Jia-YingJia-YingSuTUNG-HUNG SUSun, Hsun-IHsun-ISunHuang, Wei-MinWei-MinHuangTsai, Pei-LingPei-LingTsaiWernig, GerlindeGerlindeWernigHo, Ping-ChihPing-ChihHoWang, LimeiLimeiWangTSENG-CHENG CHENYIH-LEONG CHANGMeng, Tzu-ChingTzu-ChingMengCHEN-TU WUChang, Yao-MingYao-MingChangLai, Shih-LeiShih-LeiLaiLi, Chia-WeiChia-WeiLiKo, Tai-MingTai-MingKoChang, Ya-JenYa-JenChangKAI-CHIEN YANGChern, YijuangYijuangChernKuo, Mei-ChuanMei-ChuanKuoHuang, Yen-TsungYen-TsungHuangTzeng, Yi-ShiuanYi-ShiuanTzengJIH-LUH TANGChen, Shih-YuShih-YuChen2025-10-222025-10-222025-07-01https://scholars.lib.ntu.edu.tw/handle/123456789/732787In eukaryotic cells, reactive oxygen species (ROS) serve as crucial signaling components. ROS are potentially toxic, so constant adjustments are needed to maintain cellular health. Here we describe a single-cell, mass cytometry-based method that we call signaling network under redox stress profiling (SN-ROP) to monitor dynamic changes in redox-related pathways during redox stress. SN-ROP quantifies ROS transporters, enzymes, oxidative stress products and associated signaling pathways to provide information on cellular redox regulation. Applied to diverse cell types and conditions, SN-ROP reveals unique redox patterns and dynamics including coordinated shifts in CD8 T cells upon antigen stimulation as well as variations in CAR-T cell persistence. Furthermore, SN-ROP analysis uncovers environmental factors such as hypoxia and T cell exhaustion for influencing redox balance, and also reveals distinct features in patients on hemodialysis. Our findings thus support the use of SN-ROP to elucidate intricate redox networks and their implications in immune cell function and disease.en[SDGs]SDG3[SDGs]SDG15journal article10.1038/s41467-025-60727-z40593569