Xenbase Image ID: 145099

Figure S3. Anillin regulates the distribution of Rho-GTP, F-actin, and P-MLC, Related to Figure 4. A. Frames from live confocal imaging of Rho-GTP dynamics in dividing control and Anillin KD embryos. Embryos were injected with GFP-rGBD (Rho binding domain of Rhotekin) as a probe for active Rho. In control embryos, Rho-GTP is present around cell-cell junctions and remains focused at the equatorial cortex (red arrow) throughout cytokinesis. When Anillin is knocked down, active Rho is not restricted to the equatorial cortex (red arrow). Flares of Rho-GTP appear at cell-cell junctions around the perimeter of the cell (yellow arrows). B. F-actin (mChe-UtrCH) images corresponding to late cytokinesis in the control and Anillin KD cells in Figure S3A showing that the dividing Anillin KD cell is separating from its neighboring cells (yellow arrows), whereas the control cell maintains adhesion with its neighbors. C. Brightest point projections of Rho-GTP flares over three time intervals in dividing control and Anillin KD cells (division site marked by red arrow). Red = flare during minutes 0-2.5, green = flare during minutes 2.5-5, blue = flare during minutes 5-7.5, white = overlap of Rho-GTP flares through the time intervals. Boxed regions are enlarged in Figure S3C’. C’. Montage of frames from time lapse movies corresponding to the dashed boxes marked in the Anillin KD images in Figure S3C highlighting the lifetime of the Rho-GTP flares. D. Scheme showing how kymographs were generated for analysis of junctional RhoGTP over time (see Experimental Procedures). Tricellular junctions where three cells come together are numbered above the resulting kymographs and are marked by vertical lines in the kymographs. Image published in: Reyes CC et al. (2014) Copyright © 2014. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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