Lipid ROS assay and MDA assay

C11-BODIPY 581/591 (GC40165, Glpbio, USA) was used to measure the levels of lipid reactive oxygen species in a flow instrument. First, we stimulated the cells with different concentrations of LPS, LPS + Mel, Era (Erastin, An activator of ferroptosis) stimulants for 24 h, respectively, and then, 5-10 × 10⁵ cells were centrifuged at 2000rpm for 3 min at room temperature, and the supernatant was discarded. Each tube received 100 μl of C11-BODIPY probe at a final concentration of 2 μM and was incubated in a cell incubator for 30 min. Then, centrifuge at room temperature for 3 min at 2000rpm, discarding the supernatant. Each tube should be filled with 300 μl of PBS, centrifuged for 3 min at room temperature at 2000rpm, the supernatant discarded, and then two more times washed. 300 μl PBS was resuspended, stored in a 4°C refrigerator away from light, and detected in time by flow cytometry. Malondialdehyde (MDA) level was determined by MDA assay kit (BC0025, Solarbio, China)