Table 1.
List of antibodies.
Fig 1.
Tumour growth rate and mouse survival are unaffected by exercise in B16-F10 and EO771 tumours.
Survival curves for mice bearing B16-F10 (a) or EO771 (b) tumours (endpoint due to tumour size only, mice euthanised due to other endpoints were excluded from the survival analysis). Average tumour growth rate in mice bearing B16-F10 (c) or EO771 tumours (d). Exercising mice (Ex) received a running wheel, while non-exercising controls (No Ex) did not. Data are shown as mean ± SD. n = 3–12 for B16-F10 or n = 3–10 for EO771 (progressively fewer mice over time as they were euthanised).
Fig 2.
Running distance of tumour-bearing mice.
Average daily running distance on a running wheel by mice bearing B16-F10 (a) or EO771 (b) tumours. Data are shown as mean ± SD; n = 3–12 (progressively fewer mice over time as they were euthanised).
Table 2.
Body and organ weights of non-exercising vs exercising mice with B16-F10 or EO771 tumours.
Fig 3.
Exercise reduces variance in perfusion in EO771 tumours.
(a) Representative immunofluorescent images of sections containing Hoechst 33342 (perfused blood vessels, blue) and stained for pimonidazole (hypoxia, green) in B16-F10 and EO771 tumours from non-exercising vs exercising mice. Quantification of hypoxic area in B16-F10 (b) and EO771 (d) tumours from non-exercising vs exercising mice. Quantification of perfused blood vessels in B16-F10 (c) and EO771 (e) tumours from non-exercising vs exercising mice. B16-F10 No Ex and Ex: n = 12, EO771 No Ex: n = 9 (two mice had intra-peritoneal tumours and one tumour exhibited low-level, diffuse perfusion which could not be quantified) and Ex: n = 10 (one mouse had intra-peritoneal tumours, and one was euthanised before tumour development due to pyometra). Difference in variance analysed using the F test. Data are presented as individual data points and mean ± 95% CI.
Fig 4.
Exercise does not change CD31+ vessel density in B16-F10 or EO771 tumours.
(a) Representative immunohistochemical images of B16-F10 and EO771 tumours from exercising vs non-exercising mice. Quantification of the number of CD31+ vessels in B16-F10 (b) and EO771 (c) tumours from non-exercising vs exercising mice. B16-F10 No Ex: n = 8 (four tumours could not be quantified due to dark pigmentation), B16-F10 Ex: n = 10 (two tumours could not be quantified due to dark pigmentation), EO771 No Ex: n = 10 (two mice had intra-peritoneal tumours) and Ex: n = 10 (one mouse had intra-peritoneal tumours, and one pyometra). Data are presented as individual data points and mean ± 95% CI.
Fig 5.
Exercise does not change tumour cell proliferation in B16-F10 or EO771 tumours.
(a) Representative immunohistochemical staining for pHH3, a mitotic marker, in tumour sections from exercising vs non-exercising mice with B16-F10 melanoma or EO771 breast cancer. Quantification of the percentage of pHH3+ cells in B16-F10 (b) and EO771 (c) tumours from exercising vs non-exercising mice. B16-F10: n = 12 per group. EO771: n = 10 per group (three mice had intra-peritoneal tumours, and one pyometra). Data are shown as individual data points and mean ± 95% CI. Data analysed using a two-tailed student’s t test.
Fig 6.
EO771 tumours are more proliferative, more vascular and more hypoxic than B16-F10 tumours.
Exercising and non-exercising mice were pooled for this analysis as no difference was seen with exercise. (a) Quantification of the percentage of pHH3+ cells in B16-F10 vs EO771 tumours. B16-F10: n = 24, EO771: n = 20 (three mice had intra-peritoneal tumours, and one pyometra). (b) Comparison of the number of CD31+ vessels in B16-F10 and EO771 tumours. B16-F10: n = 18 (six tumours could not be quantified due to dark pigmentation), EO771: n = 20 (three mice had intra-peritoneal tumours, and one pyometra). Quantification of hypoxia (c) and perfusion (d) in B16-F10 vs EO771 tumours. B16-F10: n = 24, EO771: n = 20 (three mice had intra-peritoneal tumours, and one pyometra). Data analysed using two-tailed Mann-Whitney test. (Exercising and non-exercising animals were pooled for this analysis). Data analysed using two-tailed student’s t test. Data are presented as individual data points and mean ± 95% CI.