Extended Data Fig. 5: Removal of BAT ablates tumour microenvironmental changes and the mitigation of PI3K activation in cold-exposed tumours.

a, Fasting blood glucose concentrations of MMTV-PyMT tumour-bearing mice and Apc^Min/+ mice under 30 °C and 4 °C (n = 6 mice per group). b–f, Insulin tolerance test and glucose tolerance test of CRC tumour-bearing mice (b), MMTV-PyMT tumour-bearing mice (c, d) and Apc^Min/+ mice (e, f) under 30 °C and 4 °C (n = 6 mice per group in b-d and 4 mice per group in e, f). g, h, Immunofluorescence staining and quantifications of CA9⁺ hypoxic area, Ki-67⁺ proliferating cells, CD31⁺ microvessels, and Cl-Casp3⁺ apoptotic cells in sham-operated or BAT-removed CRC tumour-bearing mice under 30 °C and 4 °C (n = 5 or 10 random fields per group). Arrows and arrowheads point to positive signals. i, Schematic representation of the glycolysis pathway. j, Amounts of the glycolytic metabolites of CRC tumours under 30 °C and 4 °C (n = 4 biological samples per group). k, Quantification of LDH activity in CRC tumours under 30 °C and 4 °C (n = 6 tumours per group). l, m, Immunoblot analysis (l) and quantification (m) of non-phosphorylated and phosphorylated PI3K (p-PI3K) and AKT and phosphorylated AKT (p-AKT) in melanoma. Beta-actin was used for standardizing total protein loading levels (n = 4 biological samples per group). All scale bars, 50 μm. Data presented as mean ± s.e.m. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison tests (h) or two-sided unpaired t-test (a–f, j, k, m). NS, not significant; **P < 0.01; ***P < 0.001 (exact P values are presented in the source data).

Source data