Figure 3.

ROS produced by (A) CL-E1 (10 μM) in the absence (purple line) or presence (orange line) of singlet oxygen scavenger NaN₃ (10 mM) monitored by the fluorescence of the ROS sensor, DCFH₂ (5 μM), which produces the green, fluorescent product DCF upon oxidation. A higher fluorescence intensity was observed upon incubation with CL-E1 compared to CL-E1 in the presence of NaN₃. (B) Increasing concentrations of CL-E1 were incubated with DCFH₂ (5 μM), where a concentration-dependent increase in DCF fluorescence was observed. Analyzed by the two-tailed t-test, p-value <0.01 indicated by **. (C) To confirm that no ROS was produced by CL control compounds, each compound at a final concentration of 10 μM was incubated with DCFH₂ (5 μM). A minimal increase in the fluorescence of DCF was observed for all controls. Conditions: PBS pH 7.4 (5% DMSO) at 37 °C. λ_ex 490 nm. Measurements were performed in triplicate using independent samples.