Fig. 1: Hypoxia exacerbates osseous metastases in ES animal model.

a Design of the experiments testing the effect of tumor hypoxia triggered by femoral artery ligation (FAL) on ES metastasis. b–e SK-ES-1 xenografts: b Representative images of bone metastases detected by MRI and confirmed by histopathology (H&E) (n = 20). Scale bar: 400 μm. c Time line of metastasis from control or hypoxic xenografts. Hypoxia effects assessed by the generalized estimating equation (GEE) test. d The comparison of bone and extra-osseous metastases in control and FAL-treated animals detected at the end point of the experiment. One-tailed Mann–Whitney test. e Percentage of mice with bone metastases in control and hypoxic groups. One-sided χ² test. For panels c–e, n = 16 and 21 for control and FAL groups, respectively. f–h TC71 xenografts: f Representative image of bone metastasis detected by histological analysis (n = 7). Scale bar: 1000 μm. g The analysis of bone and extra-osseous metastases in control or FAL-treated mice. Two-tailed Mann–Whitney test. h Percentage of mice with bone metastases in control and hypoxic groups. Two-sided Fisher’s exact test. For panels g, h, n = 15 and 19 for control and FAL groups, respectively. Error bars indicate standard error of the mean.