Fig. 5. In vitro characterization of cell death induced by QN/TMZ association.

Effects of CA074 and ferroptosis inhibitors in GSC#1. a FACS analysis after staining with calcein-AM in GSC#1 untreated or treated 72 h with 450 μM TMZ, 5 μM QN or their association in the presence or absence of CA074, DFO, or both. Values in ordinate represent the percentage of dead calcein-negative cells. b GSH intracellular content after cell staining with MCB. c Mitochondrial membrane potential (MMP) analysis performed by using TMRM. d Mitochondrial ROS production by using MitoSOX-red. Bar graphs show results obtained from three independent experiments performed in duplicate and reported as means ± SD. In ordinate, the median fluorescence intensity. (*) and (**) indicate p < 0.05 and p < 0.01, respectively, vs. TMZ/QN-treated cells. (°°) indicate p < 0.01 vs. untreated samples