Fig. 4: Hypoxia correlates with pro-metastatic transcription signatures.

a Heatmap depiction of the expression of a panel of metastasis-associated genes from n = 3 Hif1α^−/− tumors compared to the average expression of n = 3 Hif1α^f/f tumors. The difference in expression of each gene between the three Hif1α^f/f and three Hif1α^−/− tumors was compared using the Mann–Whitney test. b Expression of a panel of EMT-related genes (Cdh1, Cdh2, Egfr, Notch1, Snai1, Snai2, Snai3, Twist1, Zeb1, Zeb2) compared to B2m, measured by qPCR. Two-tailed Mann–Whitney test against corresponding f/f control. c Expression of a panel of autophagy-related genes (Atg7, Becn1, Map1lc3a, Map1lc3b, Sqstm1, Ulk1) compared to B2m, measured by qPCR. Two-tailed Mann–Whitney test against corresponding f/f control. d, e Correlation of Ye et al. hypoxia gene signature⁴⁵ with mRNA levels of genes in the Kang et al. bone metastasis signature⁴⁶ or the Minn et al. lung metastasis signature⁴⁷, respectively, in TCGA Invasive Breast Carcinoma patient dataset. Pearson and Spearman correlation. n = 1100 patients. f Quantification of Ki-67⁺ area as a percentage of the lesion area. Two-tailed Mann–Whitney test. n = 17 individual Hif1α^f/f metastatic lung lesions, n = 47 individual Hif1α^−/− metastatic lung lesions. g Representative images of Ki-67-stained lung metastatic lesions, taken with ×20 objective. Scale bars represent 200 μm. h Quantification of the average number of CD4⁺ and CD8⁺ cells in the lung normalized to the total nuclear area for the image. Data points in red denote mice that had detectable lung tumor burden. Two-tailed Mann–Whitney test against corresponding f/f control. i Representative images of immunofluorescent staining of CD4 and CD8, taken with ×20 objective. Scale bars represent 200 μm. j Quantitative PCR analysis of Lox transcript compared to B2m from primary tumor homogenate RNA. Two-tailed Mann–Whitney test. Graph represents mean per group and error bars represent s.e.m. n = 19 Hif1α^f/f PyMT⁺ mice, n = 21 Hif1α^−/− PyMT⁺ mice.