Extended Data Fig. 7: HOXB13 loss induces PCa cell motility in vitro and tumor metastasis in vivo.

a. Colony formation assays were performed in LNCaP, C4-2B and PC-3 cells with HOXB13 de-regulation or indicated treatment. The data showed that androgen-dependent LNCaP cell growth was abolished by HOXB13 KD, which could be restored by re-expression of either WT or G84E HOXB13, suggesting an AR-dependent effect. By contrast, C4-2B is only partially sensitive to HOXB13 KD, whereas the growth of C4-2B cells pre-treated by enzalutamide (ENZ) and the AR-negative PC-3 cells is unaffected by HOXB13 KD. b. Cell invasion assays of control or HOXB13-KD PC-3M cells cultured in lipid-free or regular medium. Representative images are shown (left panels), and the number of invaded cells are quantified (right panel). Scale bar: 50 µm. Quantified data shown are mean ±s.d. of three representative fields from one of three (n = 3) independent experiments. P values were calculated by unpaired two-sided t-test. c. Cell migration assays of PC-3 cells with control or shHOXB13. Images shown were taken at 0 and 48 hours after a scratch was created on the cell monolayer. d. Tumor volume was measured by IVIS after two weeks of intra-prostate inoculation of PC-3M cells. Y-axis shows the normalized luciferase intensity. Statistical significance was evaluated by one-way ANOVA test (P = 0.836). e. HOXB13 de-regulation did not affect PC-3M xenograft tumor growth. Tumor volume was measured every week by IVIS live mice imaging. Y-axis shows the normalized luciferase intensity. Data shown in each time point are mean ± s.d. Statistical significance was evaluated by one-way ANOVA test (P = 0.365).