Extended Data Fig. 2: Changes of the tumour microenvironment by cold in various tumour models.

a–g, Immunofluorescence staining and quantifications of morphology (H&E), CA9⁺ hypoxic area, Ki-67⁺ proliferating cells, CD31⁺ microvessels, Cl-Casp3⁺ apoptotic cells, CD45⁺ inflammatory myeloid cells, IBA1⁺ pan-macrophages, and FSP1⁺ fibroblasts in melanoma (a, b), mouse PDAC (c, d), and human PDAC (f, g) under 30 °C and 4 °C (n = 5, 8, or 10 random fields per group). Arrows and arrowheads point to positive signals. Tumour growth of human PDAC (e) under 30 °C and 4 °C (n = 8 mice per group). h–k, Immunofluorescence staining and quantifications of CA9⁺ hypoxic area, Ki-67⁺ proliferating cells, CD31⁺ microvessels, and Cl-Casp3⁺ apoptotic cells in MMTV-PyMT tumours (h, i), and Apc^Min/+ polyps (j, k) under 30 °C and 4 °C (n = 6 or 8 random fields per group). Arrows and arrowheads point to positive signals. Scale bars in the upper panel of j, 1 mm. All other scale bars, 50 μm. Data presented as mean ± s.e.m. Statistical analysis was performed using two-sided unpaired t-test (b, d, e, g, i, k). NS, not significant.

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