Dynamic roles of p53-mediated metabolic activities in ROS-induced stress responses

Figures & data

Figure 1. Wild type p53 facilitates ferroptosis through SLC7A11 under ROS-induced stress. (A) p53^WT tet-on stable line cells were pre-treated with doxycycline (0.1 µg/mL) for 24 hours before ROS (TBH, 60 µM) was added for 3 hours with or without Ferr-1. (B) Quantification of cell death as shown in (A). (C) p53^WT tet-on stable line cells were transfected with control plasmid or plasmid overexpressing SLC7A11. Cells were seeded 24 hours later and after attachment, cells were induced by doxycycline (0.1 µg/mL) for 24 hours before addition of ROS (TBH, 30 µM) for another 3 hours. (D) Quantification of cell death as shown in (C). *, P < 0.01, n.s, not significant (Student's t test).

Figure 2. (A) Functional N-terminal domain of p53 is required to downregulate SLC7A11 and to promote ferroptosis. (A) Schematic diagram showing the locations and sequences of 2 p53 mutants, p53,^25,26,53,54 and p53^3KR. (B) H1299 tet-on p53^3KR and p53,^25,26,53,54 stable line cells were induced by doxycycline (0.1 µg/mL) and total cell lysate was analyzed by western blots for the expression of MDM2, SLC7A11, p53 and VINCULIN. (C) H1299 tet-on p53^3KR and p53,^25,26,53,54 cells were pre-treated with doxycycline (0.1 µg/mL) for 24 hours and then treated with erastin (10µM); images were taken 40 hours thereafter. (D) Quantification of cell death as shown in (C). *, P <0.01 (Student's t test).

Figure 3. p53 dynamically regulates intracellular ROS. (A) p53^3KR and p53,^25,26,53,54 tet-on stable line cells were treated with doxycycline (0.1 µg/mL) for indicated time and ROS levels were determined. (B) Comparison of functional activities among p53^WT, p53^3KR and p53,^25,26,53,54. *, P <0.01, n.s, not significant (Student's t test).