Figure 8

The cell-free supernatant from TCR stimulated CD4⁺CD25^- T cell culture contained ROS. CD4⁺CD25^- T cells were cultured with anti-CD3 and anti-CD28 for the indicated time points and ROS in the culture supernatant was detected using DCFH-DA as described in the Method section. Oxidation of DCFH-DA was measured using a spectrofluorometer at wavelength 485/535 nm and is represented as fluorescent units. The experiment was repeated twice with similar results.