Intracellular ROS decrease mediated by TAT-fused protein delivery.
(A) Strategy for reactive oxygen species quantification using DCFH-DA probe. (B) Cell monolayers were pre-treated for 2 h with 500 μM H2O2. Medium was discarded and cells were loaded with 80 μM DCFH-DA (30 min) and washed. Cells were either incubated in serum free medium alone or treated with 3 μM TAT fused proteins and 100 μM Chloroquine (1 h). CQ represents a control where DCFH-DA loaded cells were subsequently incubated with 100 μM chloroquine alone. After trypsinization, cells were resuspended in PBS buffer and transferred to a black 96 well plate to read fluorescence at 520 nm (excitation: 485 nm). n = 3. **p<0.01 vs. CON (control) cells; ***p<0.001 vs. CON (control) cells; ##p<0.01 vs. H2O2 alone treated cells.