Fig. 6.

Progesterone and estradiol synergistically promote lung metastasis of TSC2-deficient cells in a preclinical model of LAM. ELT3 cells were subcutaneously injected into female ovariectomized mice implanted with slow-releasing pellets of E₂, progesterone, E₂ + progesterone, or placebo. a The tumor volume was calculated at 8-week post-cell inoculation. b The number of lung metastases in female mice was scored: placebo (P) (n = 10), E₂ (n = 8), progesterone (Pg) (n = 4), and E₂ plus progesterone (E ₂ + Pg) (n = 4). *p < 0.05, **p < 0.01, Student’s t test. c Lung sections from female mice were stained with H&E. Arrowheads point to metastatic lesions in mouse lungs. d Immunohistochemical staining of smooth muscle actin (SMA) and phospho-S6 (Ser235/235) in mouse lung sections. e Immunofluorescent double staining of phospho-S6 and PCNA in mouse lung sections