Fig. 4.

Determination of reactive oxygen species (ROS) generated by H₂O₂ in GS28 siRNA-transfected cells. siRNA-transfected cells were treated with 200 µM H₂O₂ for 3 h in the presence of BSO without or with pretreatment with 40 µM necrostatin-1 (Nec-1), 1 mM tempol and 50 µM deferoxamine (DFO). The cells were incubated with 5 µM H₂DCF-DA, and fluorescence intensities were measured in flow cytometry, as described in Materials and Methods.