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. Author manuscript; available in PMC: 2014 Oct 14.
Published in final edited form as: Dev Cell. 2013 Sep 26;27(1):47–59. doi: 10.1016/j.devcel.2013.08.002

Figure 1. The chromosome-binding protein Dppa2 is essential for sperm pronuclear assembly in Xenopus egg extracts.

Figure 1

(A) Chromosomal localization of Dppa2 protein in interphase and metaphase visualized by immunofluorescence. Right: linescan of fluorescence intensity across the metaphase spindle axis.

(B) Dppa2 protein levels in mock-depleted (Control), Dppa2-depleted (ΔDppa2) and Dppa2-depleted extracts reconstituted with recombinant Dppa2 protein (ΔDppa2 + Dppa2) were analyzed by Western blotting.

(C) Demembranated sperm nuclei were added to metaphase extracts together with calcium to release into interphase and initiate pronuclear assembly. Samples were fixed and stained with Hoechst 33342.

(D) Quantification of nuclear cross-sectional area and roundness from (C). Roundness is defined as the ratio of minor to major axes of a best-fit ellipse. Data shown are mean and standard error from > 30 nuclei per sample in a single representative experiment.

(E) Scoring of abnormal nuclear morphology from 3 independent experiments as performed in (C). Nuclei were fixed at 60 min after calcium addition. Bars indicate mean and standard error, 200 nuclei scored per sample per experiment.

Scale bars, 10 μm.

See also Figure S1.