Lysosome injury assays

1. Acridine orange-relocation assay. Cells stained with 0,5 μg/mL AO (Molecular Probes) at 37°C for 15 minutes, washed twice, treated with 80 μM α-BSB for 3 hours, and analized by flow cytometry (FL-1 channel). 2. AO-uptake assay. Cells incubated with 80 μM α-BSB for 3 hours were stained with 0,5 μg/mL AO for 15 minutes at 37°C, washed twice and analized by flow cytometry (FL-3 channel). 3. LysoTracker green-uptake assay. Cells were incubated with 80 μM α-BSB for 1, 3 and 5 hours. Then they were washed and stained at 37°C for 1 hour with 75 nM of the acidotropic dye LTG (Molecular Probes) and analized by flow cytometry (FL-1 channel).