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. 2021 May 27;184(11):2860–2877.e22. doi: 10.1016/j.cell.2021.04.013

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© 2021 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Figure S1 — The parental genomes cluster with nucleoli at the pronuclear interface in live human zygotes, related to Figure 1

(A) Time between NEBD and cytokinesis in human zygotes that develop into blastocysts or developed abnormally.

(B) Classes of nucleolar spread (see (D)) changes during the last 3 hours before NEBD. Each spread trace was smoothed using local regression and mean subtracted. Soft-DTW and k-mean clustering resulted in 3 classes. Class 1 (magenta) contains nuclei with minor changes in nucleolar distribution. Class 2 (green) contains nuclei displaying active nucleolus compaction. Class 3 (blue) contains nuclei displaying active nucleolus decompaction. The percentage numbers indicate the fraction of each class. In total n = 316 nuclei were analyzed (158 zygotes). The colored lines show the barycenter of the classes.

(C) Classes of nucleolar distribution changes in (B) represented without mean subtraction. The colored lines show the barycenter of the classes.

(D) Schematic illustrating calculation of the nucleolar spread.

(E-G) Representative stills from time-lapse movies of zygotes with the indicated pronucleus belonging to class 1 (E), 2 (F), or 3 (G). The tracks of the nucleoli are shown for the indicated pronuclei, see also (H). Time, h:min, 00:00 is NEBD.

(H) Nucleolar spreads during the last 3 hours before NEBD for the zygotes shown in E, F, and G.

(I) Zygotes with or without pronuclei of class 3 that develop into blastocysts or developed abnormally.

(J) Number of nucleoli in zygote that develop into blastocysts or developed abnormally.

(K) Zygotes having less than 6 (< 6), 6, 7, or 8 (6 - 8), or more than 8 (> 8) nucleoli that develop into blastocysts or developed abnormally.

(L) Zygotes having a pronucleus with 6 or less (≤6) or more than 6 (> 6) nucleoli that develop into blastocysts or developed abnormally.

(M) Zygotes having a difference in nucleolar numbers between pronuclei of 2 or less (≤2) or more than 2 (> 2) that develop into blastocysts or developed abnormally.

(N) Representative immunofluorescence images of a zygote with 3 pronuclei. Magenta, DNA (DAPI). Gray, transmission.

(O) Pronuclei in live zygotes (as in Figure 1E) or fixed zygotes (as in (N)) were scored for the presence of DNA around nucleoli.

The number of analyzed zygotes (A, I, J, K, L, M) and pronuclei (F) is specified in italics. p values were calculated using unpaired two-tailed Student’s t test (A, J) and Fisher’s exact test (I, K, L, M). Scale bars, 10 μm.