Figure 9

Metastasis formation after injection of transformed cells into the mammary gland. Metastases were observed from 30 to 90 days after injection of oestrogen receptor alpha (ERα)/BMI1/TERT/MYC-transformed human mammary epithelial cells (HMECs). (a-e) Peritoneal metastases 90 days after injection. (f-j) liver metastases 30 days after injection. (a,f) normal light, (b,g) cyan fluorescent protein (CFP) fluorescence, (c,h) haematoxylin/eosin staining, (d,i) CFP immunostain (green) with 4',6-diamidino-2-phenylindole (DAPI) counterstain (blue), (e,j) keratin 14 immunostain (green) with DAPI counterstain (blue). (c-e,h-j) Matched tissue sections of peritoneal and liver metastases, respectively. Liver tissue is stained yellow in (i) and (j) by non-specific binding of secondary antibodies coupled to Alexa 488 and Alexa 568. Only the HMECs were infected with the CFP-expressing virus in this experiment.