Fig. 5.
Majority of Src-transformed MDCK cells extrude during or instead of mitosis. a The effect of inhibition of the G2/M transition or G1/S transition on extrusion of MDCK-pTR-cSrc-Y527F from a monolayer of normal MDCK cells. Inhibitors 2 mM hydroxyurea or 10 µM Ro-3306 were added with a 9-hour delay after the induction of Src-expression. At 24 h from Src induction cells were fixed, stained with phalloidin and imaged. Data are mean ± s.d. of three independent experiments (total number of cells: nCtrl = 155; nHU = 153; nRo3306 = 152). **P < 0.005 (Student’s t-test). b Time-lapse imaging over 24 h of extrusion of MDCK stably expressing both pTR-cSrc-Y527F-GFP and FUCCI. GFP-CAAX labels Src cells (green membrane) that express Cherry-hCdt1 in G1 phase (red nuclei) or mTurquoise-hGEM in S/G2/M phases (green nuclei). c Quantification of the nuclear colour change in MDCK-pTR-cSrc-Y527F-GFP/FUCCI cells during extrusion based on live-imaging. Data are mean ± s.d. of two independent experiments (total number of extruded cells: n = 63)