Jc 1 probe solution

After induction model, mice were anaesthetized with Isoflurane (Induction: 3-5%, Maintenance: 1.5-3%), and peripheral blood were collected form submaxillary vein. Blood was centrifugated at1000 g at 4° C for 10 min and serum was collected and saved at -80° C for other experiment. Blood or cell samples were collected and used to measure ROS (E004-1-1), TNF-α (H052-1), IL-6 (H007-1-1), IL-18 (H015) and IL-1β (H002) levels using ROS, TNF-α, IL-6, IL-18 and IL-1β ELISA kits (Nanjing Jiancheng Biological Engineering Institute, Nanjing, China) following the manufacturer’s instructions.
1x 105/well cells were seeded into a 96-well plate and 100 μL JC-1 probe solution (C2006, Beyotime Biotechnology) was added into every well as previously described [34 (link)]. Absorbance was measured using a fluorescent reader (Synergy H1 Microplate Reader, Bio Tek, VT, USA).

1x 10⁵/well of RAW264.7 cell was seeded into a 96-well plate and 100 μL JC-1 probe solution (C2006, Beyotime Biotechnology) was added into every well as previously described [52 (link)]. Absorbance was measured using a fluorescent reader (Synergy H1 Microplate Reader, Bio Tek, Winooski, USA).
1x 10⁵/well of RAW264.7 cell was seeded into a 96-well plate, diluted calcein-AM solution to 500 nM with solution buffer, incubated cells with for 20 min at 37° C, and added with 50 μL CoCl2 solution and incubated for 10 min. Absorbance was measured using the plate under Ex490/Em515 nm with fluorescent reader (Synergy H1 Microplate Reader, Bio Tek, Winooski, USA) as previously described [52 (link)].