Fig. 4: JNK downstream transcriptional factors regulate cells susceptibility to ferroptosis.

A Schematic diagram of JNK downstream transcriptional factors. B Overexpression of ATF2 promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. C Overexpression of NFATC1 promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. D Overexpression of NFATC3 promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. E Overexpression of SMAD4 promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. F Knockdown of ATF2 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. G Knockdown of NFATC1 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. H Knockdown of NFATC3 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. I Knockdown of SMAD4 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. J Erastin treatement promoted phosphorylation of ATF2, SMAD4, but suppressed phosphorylation of NFATC3 in HT1080 cells. HT1080 cells were treated with erastin (10 μM) as indicated, western blot images confirm the expression of the indicated proteins. K Overexpression of c-Jun suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. L Overexpression of ELK1 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. M Overexpression of HSF1 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. N Overexpression of STAT3 suppresses ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. O Knockdown of c-Jun promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. P Knockdown of ELK1 promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. Q Knockdown of HSF1 promotes ferroptosis. B16-F10 cells as indicated were treated with erastin (30 μM) for 14 h for cell death measurement, or 12 h for lipid ROS measurement. Cell death and lipid ROS were measured as described in Fig. 1A. Western blot images confirm the expression of the indicated proteins in indicated cells. R Erastin treatment induced time dependent phosphorylation of c-Jun and ELK1. HT1080 cells were treated with erastin (10 μM) as indicated, western blot images confirm the expression of the indicated proteins. Data are derived from three independent experiments, and each value represents the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, t test. (n = 3).