Mitochondrial Superoxide Imaging in LPS-Stimulated Macrophages
Macrophages grown on coverslips were pretreated with vehicle, 10 μM ATB1021, or 200 μM MitoTEMPO (Sigma‒Aldrich; SML0737) and then stimulated with 100 ng/mL LPS for 30 min or 2 h; mitochondrial superoxide staining was performed with 2.5 μM MitoSOX Red (Invitrogen; M36008) for 30 min. Cells were washed with PBS, fixed in 4% paraformaldehyde (PFA) for 15 min, and mounted with Fluoromount-GTM Mounting Medium with 4′,6-diamidino-2-phenylindole (DAPI; Invitrogen; 00–4959–52). After mounting, the cells were visualized using a confocal laser-scanning microscope.
Silwal P., Kim Y.J., Lee Y.J., Kim I.S., Jeon S.M., Roh T., Kim J.K., Lee M.J., Heo J.Y., Jo D.S., Lee S.H., Cho D.H., Kim J.M., Kwon Y.T, & Jo E.K. (2023). Chemical mimetics of the N-degron pathway alleviate systemic inflammation by activating mitophagy and immunometabolic remodeling. Experimental & Molecular Medicine, 55(2), 333-346.
Other organizations :
Chungnam National University, Seoul National University, Keimyung University, Kyungpook National University, Korea Basic Science Institute
Mounting with Fluoromount-G™ Mounting Medium with DAPI
controls
Positive control: Not specified
Negative control: Vehicle
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