Supplementary Materials

Spontaneous emergence of cell-like organization in Xenopus egg extracts

Xianrui Cheng and James E. Ferrell Jr

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S11
  • Captions for Movies S1 to S10
  • References

Images, Video, and Other Media

Movie S1
Homogenized Xenopus laevis egg extracts self-organize into cell-like compartments. Time lapse video of bright-field, tubulin, ER and tubulin-ER overlay channels of cell-like compartment formation dynamics in interphase Xenopus laevis egg extracts. Related to Fig. 1C.
Movie S2
Detailed view of spontaneous cell-like compartment formation in Xenopus laevis egg extracts. Time lapse video of bright-field, tubulin, ER and tubulin-ER overlay channels of cell-like compartment formation dynamics in interphase Xenopus laevis egg extracts, focusing on a small region of the imaging field to reveal more details. The migration of nuclei from their initial locations during the compartment formation process is readily visible, indicating that the initial localization of added demembranated sperm nuclei does not entirely determine the final pattern of the compartments. Related to Fig. 1C.
Movie S3
The organization of microtubules, endoplasmic reticulum, mitochondria and the nucleus in the cell-like compartment resembles that of a typical cell. Time lapse video of bright-field, tubulin, mitochondrial, ER, nuclear and overlay channels of cell-like compartment formation dynamics in interphase Xenopus laevis egg extracts. Related to Fig. 1D.
Movie S4
The formation of cell-like compartments in egg extracts does not require added demembranated sperm nuclei. Time lapse video of bright-field, tubulin, ER, nuclear and overlay channels of cell-like compartment formation dynamics in interphase Xenopus laevis egg extracts with added demembranated sperm nuclei (top panels) and without added nuclei (bottom panels). Related to Fig. 2A and B.
Movie S5
Confocal view of the formation of cell-like compartments with and without added demembranated sperm nuclei. Time lapse video of confocal images of SiR-tubulin, ER, and nuclei during pattern formation in extracts. In the merge panels, SiR-tubulin is green, ER is, and nuclei are blue. Images are maximum intensity projections from 5 z-slices 10 μm apart. Related to Fig. 1E and Fig. 2C.
Movie S6
Cell-like compartment patterns in extracts with different concentrations of added demembranated sperm nuclei. Time lapse video of cell-like compartment formation in interphase Xenopus laevis egg extracts supplemented with different concentrations of demembranated sperm nuclei. Related to Fig. 2D.
Movie S7
Formation of cell-like compartments requires microtubule polymerization. Time lapse video of cell-like compartment formation in interphase Xenopus laevis egg extracts without nocodazole treatment (top panels) and with 33 μM nocodazole treatment (bottom panels). Related to Fig. 3A and Table 1.
Movie S8
Effect of the cytoplasmic dynein inhibitor ciliobrevin D on the formation of cell-like compartments. Time lapse video of a normal extract (top) and an extract treated with ciliobrevin D (50 μM). Related to Fig. 3B and Table 1. Note that the microtubules initially concentrated in central asters and then flipped to the periphery at ~70 min.
Movie S9
The cell-like compartment is capable of mitotic division. Time lapse video of a sperm-supplemented cycling Xenopus laevis egg extract. Note that a cell-like compartment formed in the first interphase after meiotic exit by dynamics similar to those observed in interphase-arrested extracts. This compartment subsequently entered mitosis and gave rise to two daughter compartments. Related to Fig. 4A.
Movie S10
The cell-like compartment can perform multiple cycles of division and reorganization. Time lapse video of a sperm-supplemented cycling Xenopus laevis egg extract undergoing five successive cycles of division. Epifluorescence of mCherry-NLS (red), which accumulates in interphase nuclei, is superimposed on bright-field images.