A Protocol for Quantifying Lipid Peroxidation in Cellular Systems by F2-Isoprostane Analysis
Figure 6
The F2-isoprostane approach allows for analysis of FOXO transcription factor control of endogenous oxidative stress.
(A) GOX-induced lipid peroxidation is significantly decreased when FOXO3a is activated. DLD-1 and DL-23 cells were treated with 4-OHT to activate FOXO3a for 16 hours before H2O2 exposure by GOX treatment for 2 hours. F2-isoprostane levels were analyzed by UPLC-MS/MS and are shown as mean fold increase ± S.D (* p < 0.05) (B) Prolonged FOXO3a activation increased the protection against endogenously formed ROS damage in colon carcinoma cells. DLD-1 and DL-23 cells were incubated with 4-OHT to activate FOXO3a for 24 hours, before treatment with PQ for another 8 hours. 5-iPF2α-VI levels were analyzed and data are represented as mean fold increase ± S.E.M (*p < 0.05, *** p < 0.001).