Fig. 3
Molecular changes associated with the collective extrusion. a Confocal image of F-actin after 60 h of blue-light illumination. Maximum intensity projection of a Z-stack of 32 slices with 1 μm slice interval with orthogonal views from the confocal stack. The blue circle delineates the region of blue-light illumination. (r,Ɵ) are the polar coordinates used for averaging in (b). b Average radial profile of the gray value of the image depicted in (a). The blue shaded area indicates the region of stimulation. c Effect of drugs modifying contractility on the probability of budding Pbud and the appearance time Tap. Calyculin A (1 nM) decreases Tap (p = 0.021, Wilcoxon rank sum test) and increases Pbud, while Y27632 (10 μM) increases Tap (p = 0.013, Wilcoxon rank sum test) and decreases Pbud. Horizontal black lines correspond to the mean values. The error bar of Pbud is the uncertainty as described in the Methods (nCal = 28, nDMSO = 30, nY27632 = 55 and nwater = 24, all coming from at least 3 different experiments). d, e A monolayer of Madin-Darby Canine Kidney (MDCK) optoSrc cells was subjected to the standard conditions of illumination for 60 h and fixed with paraformaldehyde. d Immunostaining for E-cadherin (orange) in a monolayer displaying a budding aggregate reveals that E-cadherin is depleted from the membrane for Src-activated cells (black circle) compared to non-illuminated cells (dashed white square). Cell nuclei were stained with Hoechst (purple). e Reconstructed side view of a confocal acquisition after an immunostaining for vimentin (orange) in a monolayer of MDCK optoSrc cells displaying a budding aggregate. Vimentin is strongly expressed in the cells at the top of the budding structure, compared to cells in the monolayer. Cell nuclei were stained with Hoechst (cyan). Scale bars: 50 μm
