Fig. 5.

Progesterone and estradiol synergistically attenuate the death of TSC2-deficient cells under oxidative stress. a Patient-derived TSC2-deficient cells or b rat-derived ELT3 cells were treated with 10 nM E₂, 10 nM progesterone (Pg), 10 nM E₂ + 10 nM Pg, or vehicle control for 24 h and then incubated with 0.5 μM H₂O₂ for 30 min. Cell morphology was recorded using phase-contrast microscopy. c Rat-derived cells and d patient-derived cells were treated with 10 nM E₂, 10 nM Pg, 10 nM E₂ + 10 nM Pg, or vehicle control for 24 h and then incubated with 0.5 μM H₂O₂ for 30 min. Cell death was measured using the propidium iodide (PI) exclusion assay. Proportion of dead cells was normalized to the total number of variable cells. Results are representative of eight independent samples per group from three experiments. e Patient-derived TSC2-deficient cells were pre-treated with ICI 182,780 (ICI, 10 μM) or RU-486 (RU, 20 μM) for 12 h, stimulated with 10 nM E₂, 10 nM Pg, 10 nM E₂ + 10 nM Pg, or vehicle control for 24 h, and then incubated with 0.5 μM H₂O₂ for 0.5 h. Cell death was measured using the PI exclusion assay. Proportion of dead cells was normalized to the total number of variable cells. Results are representative of eight independent samples per group from three experiments.*p < 0.05, **p < 0.01, Student’s t test