2.10. Evaluation of lipid peroxidation

To ascertain the involvement of lipid peroxidation in lung tumorigenesis, the cells were stained by 5 μM fluorescent probe C11-BODIPY^581/591 for 30 min at 37 °C followed by flow cytometry. To visualize the membrane, tissue slides were incubated with 25 μg/ml Concanavalin A-Alexa Fluor^TM 350 (Thermo, #{"type":"entrez-nucleotide","attrs":{"text":"C11254","term_id":"1536325"}}C11254) following 5 μM C11-BODIPY^581/591 staining at 37 °C for 30 min. Images were captured at emission at 580/600 nm (the non-oxidized form, red) and 490/510 nm (the oxidized form, green) and then merged to demonstrate the fraction of the oxidized C11-BODIPY^581/591. The malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) concentration were assessed using lipid peroxidation assay kits (Abcam, #ab118970 and #ab238538). IHC was measured to detect the expression level of 4-HNE using anti-4-HNE antibodies (Abcam, #ab46545).