Fig. 3: TRF attenuates E0771 mammary tumor growth and metastasis to lung in obese OVX mice. | Nature Communications

Fig. 3: TRF attenuates E0771 mammary tumor growth and metastasis to lung in obese OVX mice.

From: Time-restricted feeding normalizes hyperinsulinemia to inhibit breast cancer in obese postmenopausal mouse models

Fig. 3

a E0771 tumor volume over time in OVX mice on NC, AL, and TRF (number of mice n = 7 for NC, n = 7 for AL, n = 9 for TRF; ****p < 0.0001 by two-way ANOVA with Tukey’s multiple comparisons test). b Individual tumor weight at sacrifice (number of tumors n = 22 for NC, n = 28 for AL, n = 34 for TRF). ****p < 0.0001 by one-way ANOVA with Tukey’s multiple comparisons test. c Correlation plot of mean tumor weight per mouse vs. fasting insulin (number of mice n = 7 for NC, n = 7 for AL, n = 7 for TRF). Data were analyzed by linear regression using Pearson correlation. d Representative H&E, Ki67, and CD31 staining of E0771 tumor tissue in OVX mice on NC, AL, and TRF. Scale bar, 50 µm. Insets in H&E images show higher magnification of a mitotic cell. Data shown are representative of two independent experiments. e–g Quantification of mitoses (n = 20 sections for all groups), Ki67-positive cells (n = 17 for NC, n = 15 for AL, n = 16 for TRF), and CD31-positive cells (n = 12 for NC, n = 13 for AL, n = 14 for TRF) in sections of E0771 tumor tissue. Data are presented as mean ± SEM per high-powered field (hpf) or 5 hpf for mitoses, with 4 hpf per section (n = number of tumors). Asterisks indicate significance; *p < 0.05, **p < 0.01 by one-way ANOVA with Tukey’s multiple comparisons test. h Carmine Red staining of whole-mount lungs showing macro-metastases of E0771 tumor cells. i H&E stained lung sections showing metastases of E0771 tumor cells. Data shown are representative of two independent experiments. j Total number of metastases per mouse (number of mice n = 3 for NC, n = 6 for AL, n = 6 for TRF; *p < 0.05 by one-way ANOVA with Tukey’s multiple comparisons test). k Cross-sectional area of individual lung metastases (number of mice n = 3 for NC, n = 6 for AL, n = 6 for TRF; two-tailed t test). Data are presented as mean ± SEM. For a, c, h, and j, n represents number of mice per group. Asterisks show statistical significance TRF vs AL or as shown.

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