Fig. 2.

TBCP-1 tumours metastasise avidly to the brain and other organs. a Summary of the incidence of mice developing experimental or spontaneous metastases after intra-cardiac or orthotopic inoculation of TBCP-1 cells. b Quantitation of tumour burden and number of metastatic lesions/organ from experimental and spontaneous metastasis assays. Experimental femur and spine metastatic burden was quantitated by genomic qPCR of the mCherry reporter gene as described in the “Methods” section. Each dot represents one mouse, and data show mean ± SEM, n = 10. The number of lesions was determined by visual examination of histological sections. Each dot represents one mouse, and data show mean ± SD, n = 10. Representative images of soft organ metastases (red, with brightfield overlay) that developed after orthotopic inoculation of TBCP-1 cells are shown in the bottom panels. Lesions are delineated by a dotted line. c ERα, PR and HER2 were detected in TBCP-1 primary tumours or brain metastases by standard immunohistochemistry as indicated. Nuclear ERα and PR detected in normal mammary gland surrounded by a 67NR primary tumour and membrane HER2 detected in a SKBR3 primary tumour were used as positive controls (third row). H&E staining (arrows = blood vessels), pan-cytokeratin and Ki-67 positivity in TBCP-1 brain metastases are shown (bottom panels). Dotted lines delineate metastatic lesions. Scale bars = 100 μm. d Quantitation of ERα, PR, HER2, Ki-67 and pan-cytokeratin protein levels in primary tumours and brain metastases. Scoring of tissue sections was done using the ImageScope software as described in the “Methods” section. Data show mean ± SEM of three independent tumours/metastases