Fig. 4

Re-adhesion reveals that active Ras promotes abscission in the detached cells. Mitotic BJ-LT and BJ-LT-Ras cells were isolated and either seeded directly on fibronectin (control) or cultured in suspension for 3 h before re-plating on fibronectin-coated coverslips. a Bright field micrographs from representative time-lapse movies showing the separation of two daughter BJ-LT-Ras cells at the indicated time points during re-adhesion to fibronectin directly (upper panel) or after 3 h suspension culture (lower panel); the latter cell is larger because of growth during the 3-h period. b, c Representative immunofluorescence micrographs illustrating the distribution of septin-7 (green) and α-tubulin (red) in BJ-LT-Ras (b) and BJ-LT after re-plating on fibronectin for 40 min with a pre-incubation for 3 h in suspension. Nuclei were stained with DAPI (blue). c % of cells that were separated from each other after 40 min re-plating as shown in b and after adhesion during the entire period of 3 h + 40 min (control cells)