Fig. 3
Cells that detach collectively have a survival advantage over detaching single cells and are proliferative. a DIC/fluorescence images from representative time-lapse series of single and cluster detachment for OV90 and OVCAR3 cells. Cells were treated with 2 µM CellEventTM Red dye (Invitrogen), a fluorescent marker for active Caspases 3 and 7. b Quantification of apoptosis events in single cells and clusters during detachment from organoids in OV90 (n = 47; p < 0.001) and OVCAR3 (n = 23; p < 0.001) cells. Some clusters were predominantly alive, but contained at a single dead cell, and were labeled as “Both”. c Immunofluorescent images of primary, ascites, and peritoneal wall metastases stained for the proliferation marker Ki67 and counter-stained for E-cadherin. Paraffin-embedded tissue was processed by the GCRC Histology Core Facility, and antigen-retrieval and immunostaining of tissue sections were performed as described previously [30]. Antibodies and concentrations are listed in Supplementary Table 1. d Quantification of Ki67 positive cells from the indicated sites; n = 3; p > 0.05; error bars, s.d.
