Extended Data Fig. 9: The PFKFB4–SRC-3 axis promotes breast tumour growth and metastasis.
From: Metabolic enzyme PFKFB4 activates transcriptional coactivator SRC-3 to drive breast cancer
a, Primary tumours resected out after 6 weeks. b, Ki67 staining of primary tumours from animals injected with MDA-MB-231 cells stably expressing control shRNA, SRC-3 shRNA or PFKFB4 shRNA. Data are representative of five fields per slide from n = 5 animals per group with similar findings. Scale bar, 100 μm. c, Quantification of Ki67-positive cells in the tumour. n = 5 animals per group, average of five fields counted from each slide. ****P = 0.0001, one-way ANOVA with Dunnett’s multiple comparisons test. d, Primary tumour growth in animals injected with MDA-MB-231 cells stably expressing shRNA targeting SRC-3, PFKFB4, or expression of wild-type SRC-3 or the Ser857Ala mutant in the SRC-3-depleted cells. n = 5 animals per group. *P < 0.000001, two-way ANOVA with Tukey’s multiple comparisons test. e, Immunoblot showing the relative expression of SRC-3 in primary tumours from MDA-MB-231 cells stably expressing control shRNA, SRC-3 shRNA, or after re-expression of wild-type SRC-3 or the Ser857Ala mutant in the SRC-3-depleted cells. n = 5 animals per group was pooled to generate the tumour lysate used for analysis. f, Graph representing the photon flux of animals from different groups. n = 5 animals for wild-type SRC-3, the Ser857Ala mutant and PFKFB4 shRNA, and n = 4 animals for SRC-3 shRNA. *P < 0.0001, one-way ANOVA with Dunnett’s multiple comparisons test. Line shows median with range. g, mRNA expression of three metabolic enzymes (TKT, XDH and AMPD1), SRC-3 and PFKFB4 from the primary tumours. n = 5 animals per group. **P < 0.05, ***P < 0.001, ****P = 0.0001, two-way ANOVA with Tukey’s multiple comparisons test. h, Expression of PFKFB4 in patients with breast cancer across different subtypes compared to normal breast tissue. Normal basal = 17; basal = 139; normal_Her2 = 9; Her2 = 67; normal luminal A = 62; luminal A = 418; normal luminal B = 21 and LumB = 186. Line in the centre of the rectangle represents the median, top edge of the rectangle represents the third quartile, bottom edge of the rectangle represents the first quartile, top whisker represents the maximum and bottom whisker represents the minimum. All data are representative of three biologically independent experiments with similar results, and are shown as mean ± s.d. unless otherwise stated. See Source Data for exact P values.
