Supplemental Materials
This article contains the following supporting material:
- Supplemental Materials
- Movie01 - Supplemental Video 1. Entotic vacuole undergoes fission. Entotic vacuole harboring H2B-mCherry-expressing cell corpse undergoes fission as corpse is degraded. Video shows confocal time-lapse analysis of MCF10A cells. Left image panel shows DIC from a single x-y plane, right image panel shows mCherry fluorescence (red) as a maximum projection. Acquisition times are shown as minutes (min). Note engulfing cell also expresses H2B-mCherry and is binucleate.
- Movie02 - Supplemental Video 2. Apoptotic cell phagosome undergoes fission. Apoptotic cell phagosome undergoes fission as corpse is degraded. Video shows confocal time-lapse analysis of J774.1 macrophage phagocytosing an apoptotic cell expressing H2B-mCherry. Left image panel shows DIC and mCherry fluorescence (red) from a single x-y plane, right image panel shows mCherry fluorescence (red) as a maximum projection. White outline shows area of macrophage; video frames were aligned by tracking the movement of the engulfing cell. Times are shown as minutes (min).
- Movie03 - Supplemental Video 3. Entotic vacuole undergoes fission into lysosome network. Video shows confocal time-lapse analysis of Lamp1-GFP-expressing MCF10A cell (green) harboring an entotic vacuole with an H2B-mCherryexpressing corpse (red). Times are shown as minutes (min). As entotic vacuole undergoes fission, the Lamp1-GFP-labeled lysosomes in the engulfing cell become labeled with mCherry.
- Movie04 - Supplemental Video 4. Lysosomes fuse to entotic vacuoles. Two control entotic vacuoles containing cell corpses (arrows) at different stages of degradation (corpse 1, top; corpse 2, bottom) fuse with Lamp1-GFP-labeled lysosomes (green) after cell fusion initiated by PEG treatment. Arrows follow entotic vacuole in merged image. Time indicates minutes after cell fusion. 10 out of 10 entotic vacuoles in control cells recruited Lamp1-GFP within 3 hours after cell fusion.
- Movie05 - Supplemental Video 5. Lysosomes fuse to entotic vacuoles in Brefeldin Atreated cells. Two entotic vacuoles in containing cell corpses (arrows) in Brefeldin A-treated cells (corpse 1, top; corpse 2, bottom) fuse with Lamp1-GFP-labeled lysosomes (green) after cell fusion initiated by PEG treatment. Arrows follow entotic vacuole in merged image. Time indicates minutes after cell fusion. 10 out of 10 entotic vacuoles in Brefeldin A-treated cells recruited Lamp1-GFP within 3 hours after cell fusion.
- Movie06 - Supplemental Video 6. Entotic vacuole exhibits tubulation. Video shows confocal time-lapse analysis of entotic vacuole in Lamp1-GFP-expressing cell (green) harboring an H2B-mCherry-expressing corpse (red). Images show maximum projections of merged green and red fluorescence acquired every 5.5 seconds through the entire z-stack covering the vacuole, at 0.5 micron steps. Time is shown as seconds (sec).
- Movie07 - Supplemental Video 7. Entotic corpse degradation and vacuole shrinkage. Video shows entotic corpse expressing H2B-mCherry and entotic vacuole shrinking in a control MCF10A culture. Left image panel shows merged DIC and mCherry fluorescence (red), right image panel shows mCherry fluorescence (red). Times are shown as hours (hr).
- Movie08 - Supplemental Video 8. mTOR inhibition slows entotic vacuole shrinkage. Video shows entotic corpse expressing H2B-mCherry and entotic vacuole shrinking in a Torin1-treated MCF10A culture. Left image panel shows merged DIC and mCherry fluorescence (red), right image panel shows mCherry fluorescence (red). Times are shown as hours (hr).
- Movie09 - Supplemental Video 9. Lysosomes fuse to entotic vacuoles in Torin1-treated cells. Two entotic vacuoles in containing cell corpses (arrows) in Torin1-treated cells (corpse 1, top; corpse 2, bottom) fuse with Lamp1-GFP-labeled lysosomes (green) after cell fusion initiated by PEG treatment. Arrows follow entotic vacuole in merged image. Time indicates minutes after cell fusion. 10 out of 10 entotic vacuoles in Torin1-treated cells recruited Lamp1-GFP within 3 hours after cell fusion.