The real-time GSH dynamics in living BMDMs were determined with the fluorescent GSH probe, RealThiol RT-AM as described37 (link). Briefly, cells were plated on 2-well chamber Nunc slide pre-coated with 25 μg/ml fibrin with or without 5 μM acivicin or GGsTop. Buthionine sulfoximine (BSO, Sigma-Aldrich) was used as positive control for reducing intracellular GSH levels at 100 μM. Cells were kept at 37 °C during the entire experiment. After 24 h incubation, cells were loaded with 1 μM RT-AM probe for 5 min then fluorescence emissions after sequential excitation at 405 nm and 488 nm was acquired using sequential confocal laser scanning microscopy (Olympus FV1000; Olympus) with a 10× objective and 2× optical magnification. For each independent experiment, laser power and detector settings were set using the control/untreated cells incubated with RT-AM alone and then all settings were kept constant throughout the experiment. The ratio bound intracellular RT-AM:unbound intracellular RT-AM for each treatment condition was calculated by subtracting the 488-nm fluorescence signal from the 405 nm fluorescence signal from 30–50 cells/treatment for each independent experiment. The ratio calculated for each treatment was expressed as percentage from that of the untreated cells (control).