Fig. 3
Notch2HIGH breast cancer cells. a Size, granularity and b percentage of MDALUC Notch2HIGH cell subpopulation after staining with anti-Notch2 antibody, analysed by flow cytometry. c Percentage of Notch2HIGH MDALUC cell subpopulation measured after magnetic-activated cell sorting (MACS). d Notch2HIGH MDALUC cells analysed in overt osteolytic breast cancer bone metastases. White arrowhead, Notch2HIGH cell. e Triple immunofluorescence in the tibias of mice injected into the medullary cavity with breast cancer 4T1 cells. f Quadruple immunofluorescence in the tibias of the mouse model of MDA-MB-231 cell dormancy and g in the mouse model of chemotherapy-induced dormancy. B bone. h Graphical scheme of MACS-sorted Notch2HIGH and Notch2LOW MDALUC injection in 5-week-old female immunocompromised CD1 nu/nu mice. i Mouse body weight measured 6 weeks after cancer cell injection. j Micro- computed tomographic analysis of bone volume over total tissue volume (BV/TV) in tibias injected with tumour cells. k Immunofluorescent human cytokeratin area over tissue area, l percentage of metastatic area over total tissue area and m number of multicellular metastases per mm2 in the histological sections of livers collected from the mice intratibially injected with tumour cells shown in i and j. Imaging and data (mean ± SD) represent the results of at least four mice/group or three independent in vitro experiments. Statistical analyses: two tails’ unpaired t test
