bioimagesUnknownGloria SlattumThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-200903079JCB

S Movie 2. An apoptotic cell extruding basally from an MDCK cell monolayer. Note that the cell that extrudes basally (blue arrow in beginning frame) appears to migrate under the monolayer, and presumably is later engulfed. Frames were taken every 60 secs using a 40X Nikon phase lens 0.75.

Myosin IIA and phospho-myosin II are basolateral during apical extrusion and apical during basal extrusion. Note that the time dimension here actually represents different Z slices from basal to apical.

Treatment with nocodazole disrupts basolateral distribution of p115 RhoGEF (red and middle panels) and RhoA (green and bottom panels), where white arrows point to dying cell (DNA is blue). Note that the time dimension here actually represents different Z slices from apical to basal.

EB1 (green), a plus-end microtubule binding protein, colocalizes with the myosin II (red) ring during apical extrusion. Note that most EB1 is from 0µm-4µm from the base during apical extrusion. Note that the time dimension here actually represents different Z slices from basal to apical.

EB1 (green), a plus-end microtubule binding protein, colocalizes with the myosin II (red) ring during basal extrusion. Note that the time dimension here actually represents different Z slices from basal to apical.

p115 RhoGEF (red, and separate middle panel) form filaments that point towards the basolateral surface of the extruding ring during apical extrusion, while RhoA (green and bottom panel) is present through apical and basal regions of the ring. Note that the time dimension here actually represents different Z slices from basal to apical.

p115 RhoGEF and microtubules colocalize near extrusion ring. Treatment with taxol disrupts basolateral distribution of p115 RhoGEF (red and middle panels) and RhoA (green and bottom panels), where white arrows point to dying cell (DNA is blue). Note that the time dimension here actually represents different Z slices from apical to basal.

Microtubules (green) target towards the top of the extruding actin ring (red) throughout the extrusion process. Note that far fewer microtubules target than in the apical extrusion case (S Movie 8). Movie was acquired on an inverted Nikon with a 60x lens using a Yokagawa spinning disc and Andor 1000 cooled ccd camera. Four Z-section planes were taken through 9µm and projected together using 4-D software from Andor. Entire movie represents one hour.

S Movie 1. An apoptotic cell extruding apically from an MDCK cell monolayer. The cell that will extrude is marked by a blue arrow in the beginning frame. Frames were taken every 60 secs using a 40X Nikon phase lens 0.75.

Note that the cell that extrudes basally (blue arrow in beginning frame) appears to migrate under the monolayer, and presumably is later engulfed. Frames were taken every 60 secs using a 40X Nikon phase lens 0.75. Note that the Z dimension here actually represents a time lapse series.

S Movie 5. Basal extrusion of a dying cell in the epidermis of a 4 day old zebrafish treated with taxol. Dying cell extrudes basally into the epidermis and is trapped beneath the epidermis when zebrafish larvae are treated with 450µg/ml G-418 and 20 µM taxol. Frames were taken every 60 secs using a 40X Nikon water immersion DIC lens 0.8W. Note that the Z dimension here actually represents a time-lapse series.

Microtubules (green) target towards the extruding actin ring (red) early during the extrusion process and remain throughout until the ring has closed. Then the microtubules retract from this point. Movie was acquired on an inverted Nikon with a 60x lens using a Yokagawa spinning disc and Andor 1000 cooled ccd camera. Four Z-section planes were taken through 9 µm and projected together using 4-D software from Andor. Entire movie represents one hour.

S Movie 4. Apical extrusion of a dying cell from the epidermis of a 4 day old zebrafish. Dying cell extrudes apically out of the epidermis and is trapped between fish and coverglass when zebrafish 4 day old larvae are treated with 450µg/ml G-418 and 0.2% DMSO (as carrier control). Frames were taken every 60 secs using a 40X Nikon water immersion DIC lens 0.8W. Note that the Z dimension here actually represents a time-lapse series.

Treatment of 4-day old zebrafish larvae with G-418 to induce apoptosis and with taxol to stabilize microtubules drives extrusion basally. Note that the actin (red) ring are at the top (apex) of fish and the condensed DNA (blue) is basal, where numbers represent µm from most apical part of ring. At this stage, active caspase-3 (green) is dim, but will become brighter later as cell extrudes (as seen in supplementary movie 7).

bioimagesFig1video1Image 4699 (Fig1video1 - None)Image 4700 (Fig1video2 - None)Image 4705 (Figure 3 - B)Fig1video5Image 4703 (Figure 2 - c)Image 4706 (Figure 5 - A)Fig1video4Image 4702 (Fig1video5 - None)Image 4707 (SFigure 2 - None)Fig1video3Fig1video2Video9Video8Figure 3 - AImage 5119 (Figure 3 - A)Figure 3 - BFig1video3 - NoneFig1video4 - NoneFig1video5 - NoneFig1video2 - NoneSFigure 3 - DFig1video1 - NoneImage 4708 (SFigure 3 - D)Figure 2 - cSFigure 3 - EImage 4701 (Fig1video4 - None)Image 4709 (SFigure 3 - E)Image 5121 (Video9 - None)Figure 5 - AImage 4710 (Fig1video3 - None)Figure 5SFigure 2SFigure 3Video9 - NoneVideo8 - NoneImage 5120 (Video8 - None)Figure 3Figure 2SFigure 2 - NoneJody RosenblattGloria SlattumKaren M. McGeefalseP115 RhoGEF and microtubules decide the direction apoptotic cells extrude from an epithelium2009-08-31T11:17:43Z2018-11-29T11:17:43Z2018-11-29T11:17:43ZS-JCBD-20090307910.1083/jcb.200903079