Fig. 5: Zingerone regulates ferroptosis sensitivity in RAW264.7 cells via p53.

A CCK-8 assay assay of RAW264.7 cells treated with different concentrations of zingerone (n = 3 independent biological replicates). B Cell viability of RAW264.7 cells treated with different concentrations of H₂O₂ was assessed using the CCK-8 assay (n = 3 independent biological replicates). C Western blot analysis verifying the effects of different concentrations of H₂O₂ on the expression of MMP9 and CTSK in RAW264.7 cells (n = 3 independent biological replicates). D Quantification of MMP9 and CTSK protein expression from (C). E Western blot analysis verifying the effects of different concentrations of H₂O₂ on the expression of p53, SAT1, and GPX4 in RAW264.7 cells (n = 3 independent biological replicates). F Quantification of p53, SAT1, and GPX4 protein expression from (E). G Knockdown of p53 with siRNA reduces ferroptosis sensitivity after zingerone treatment (n = 3 independent biological replicates). H Measurement of ROS levels in the cytoplasm before and after zingerone and p53 siRNA treatment (n = 3 independent biological replicates). Scale bar: 50 μm. I Average fluorescence intensity of ROS. J Quantification of JC-1 monomers and aggregates in RAW264.7 cells treated with or without zingerone (n = 3 independent biological replicates). Scale bar: 5 μm. K Fluorescence intensity of JC-1 aggregates/ JC-1 monomers in each group. L MDA levels. M Intracellular FeRhoNox-1 staining in cells transfected for 48 h (n = 3 independent biological replicates). Scale bar: 100 μm. N Observation of cell morphology by transmission electron microscopy after 48 h of treatment (n = 3 independent biological replicates). Scale bar: 1000 μm. Scale bar: 50 μm. The data are presented as the mean ± standard deviation. *p < 0.05, **p < 0.01, and ***p < 0.001 compared to the model group; ###p < 0.001, ##p < 0.01 and #p < 0.05 compared to the control group; ^p < 0.05 and ^^p < 0.01 compared to the treatment group.