Figure 6

AZD4547 blocks stemness of primary MECs from MMTV-ErbB2 mice. (a) Isolated primary MECs were plated for 12 days and stained to determine the colony-forming capacity of the cells. The number of colonies formed from control and AZD4547 (2 or 6 mg/kg/day)-treated mice are displayed. (b) Primary MECs were incubated for 7 days to form primary mammospheres. Primary mammospheres were counted and imaged before harvesting. A single cell suspension of primary mammosphere cells were replated for 7 days to allow secondary sphere formation. The mammosphere efficiency (number of mammospheres formed/1000 cells plated) of MECs from control and AZD4547 (2 or 6 mg/kg/day)-treated mice was recorded. (c) Isolated MECs were cultured for 7 days in matrigel for a 3D culture assay. The average numbers of colonies formed were recorded. All values are presented as the mean ± S.E. (^* P ≤ 0.05, ^** P ≤ 0.01). (d) Representative images of colonies and mammospheres from the indicated assays are shown.