Fig. 3

Thrombin induces neuronal ferroptosis. a Thrombin cytotoxicity in N27 cells. Data are means ± SEM, n = 6 wells from one representative of 3 independent experiments. b AA content was assayed from the N27 cells 24 h after thrombin (0.5 U/mL) treatment. Data are means ± SEM, n = 5 wells from one representative of 3 independent experiments. t test was performed. c Lipid ROS in N27 cells treated with thrombin for 24 h (representative histogram plot for fluorescence of oxidized BODIPY-C11). d Relative lipid ROS is expressed as the ratio of oxidized to reduced BODIPY-C11 mean fluorescence intensity in N27 cells treated with thrombin for 24 h. Data are means ± SEM, n = 3 wells from one representative of 3 independent experiments. t test was performed. e Transmission electron microscopy (TEM) of N27 cells treated with thrombin (0.5 U/mL) for 24 h. Yellow arrows indicate shrunken mitochondria. f Detection of intracellular Fe²⁺ in N27 cells treated with thrombin (0.5 U/mL) for 6 h using FerroOrange. Data are means ± SEM, n = 3 wells from one representative of 3 independent experiments. t test was performed. g Cell viability of N27 cells 24 h after thrombin (0.5 U/mL) and Liproxstatin-1 (Lip-1) of concentration gradient co-treatment. Data are means ± SEM, n = 6 wells from one representative of 3 independent experiments. h, i Cell viability of N27 cells 24 h after thrombin (0.5 U/mL), with NAC (h) or Darapladib (i) co-treatment. Data are means ± SEM, n = 6 wells from one representative of 3 independent experiments. j, k Cell viability of MDA-MB-231 cells 24 h after thrombin (0.5 U/mL), with Fer-1 (j), Lip-1 (k) co-treatment. Data are means ± SEM, n = 5 wells from one representative of 3 independent experiments