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. 2018 Apr 1;29(7):852–868. doi: 10.1091/mbc.E17-06-0374

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© 2018 Penfield et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 4: — Repair machinery is recruited to NE rupture sites marked by Nup160 gaps and LEM-2 foci at the nuclear rim. (A) Representative examples of fixed wild-type (Ctrl) and lamin RNAi–depleted two-cell stage embryos expressing Nup160::GFP immunostained against GFP (green) and VPS-32 (red). Magnified images are of posterior (P1) nucleus. Scale bars, 5 μm. Arrowheads mark Nup160::GFP gap and VPS-32 punctum. Plots (right) of fluorescence intensities of Nup160::GFP (green) and VPS-32 (red) along three-pixel-wide lines drawn across nuclei from images shown (left). (B) Plot of percent nuclei with a visible gap at nuclear rim in GFP channel in fixed early embryos (one-cell to eight-cell) under indicated conditions. N = number of experimental repeats. (C) Select images of sperm-derived pronucleus from time-lapse series. Arrowheads mark GFP::LEM-2 and mCherry::H2B foci. Time is in seconds relative to PC regression. Percentage of nuclei containing a focus of GFP::LEM-2 at the nuclear rim is shown. Scale bar, 2.5 μm. (D) Representative examples of fixed one-cell-stage embryos expressing GFP::LEM-2 immunostained against GFP (green) and VPS-32 (red). Magnified images are of sperm-derived pronucleus. Scale bars, 5 μm. Plots (right) represent fluorescence intensities of GFP::LEM-2 (green) and VPS-32 (red) along three-pixel-wide lines drawn across nuclei from images shown (left). (E) Plot of percent nuclei with visible foci at nuclear rim. (F) Select magnified images of sperm-derived pronuclei in control and lamin-N209K–expressing one-cell stage embryos expressing GFP::LEM-2 immunostained against lamin (LMN-1, red). Scale bar, 2.5 μm. Plots (right) represent fluorescence intensities of GFP::LEM-2 (green) and lamin (red) along three-pixel-wide lines drawn across nuclei from images shown. Note different scales for LMN-1 under different conditions. (G) Plot of fold change in mean fluorescence intensity values of GFP::LEM-2 and LMN-1 at NE repair site relative to an adjacent site. n = number of nuclei. N = experimental repeats.