Fig. 4: Lamin-A regulates the F-actin meshwork, which serves as a scaffold to stabilize cytoplasmic Amot. | Nature Communications

Fig. 4: Lamin-A regulates the F-actin meshwork, which serves as a scaffold to stabilize cytoplasmic Amot.

From: The nuclear lamina couples mechanical forces to cell fate in the preimplantation embryo via actin organization

Fig. 4

a, Amot levels are increased in the cytoplasm of inner cells. Immunofluorescence images of Amot in fixed mouse embryos with standardized Amot intensity, n = 18 for outer, n = 5 for inner **P = 0.0011, Mann-Whitney U test. b, Knockdown of Lamin-A by siRNA causes an increase in cytoplasmic levels of Amot in outer cells. siRNAs were microinjected at the 1-cell stage. n = 18 for control, n = 10 for Lamin-A siRNA **P = 0.003, Mann-Whitney U test. c, F-actin meshwork density increases in the cytoplasm of inner cells at the 16-cell stage and in the ICM at the blastocyst stage. Insets show detail of cytoplasmic meshwork (cyt) and absence of actin in the nucleus (nuc). n = 5 for 16-cell inner, n = 6 for 16-cell outer, n = 8 for blastocyst ICM, n = 25 for blastocyst trophectoderm **P = 0.0087, ***P = 0.001, Mann-Whitney U test. d, Latrunculin A causes Amot disruption. Note the decrease in cytoplasmic Amot in the inner cell cytoplasm. n = 9 for control, n = 10 Latrunculin A, **P = 0.0061 Mann-Whitney U test. e, Knockdown of Lamin-A by siRNA causes an increase in cytoplasmic F-actin levels. siRNA was microinjected into a single cell of the 2-cell embryo. H2B-GFP labels injected cells. Inset shows detailed view of cytoplasmic F-actin. n = 12 for control, n = 10 for Lamin-A siRNA. ****P < 0.0001, Mann-Whitney U test. f, g, Human blastocysts immunostained for Amot (f) and stained with phalloidin (g) show increased levels of cytoplasmic Amot and F-actin in the ICM compared to trophectoderm (TE). For Amot (f), n = 2 human embryos (no statistical comparison performed). For phalloidin (g), n = 3 human embryos. P = 0.0025 by paired two-tailed t-test. For human embryos, dots represent individual cells. Fluorescence intensities represent results standardized to DAPI. Bars in dot plots represent median and interquartile range. All statistical tests are two-tailed. Scale bars, 10 µm. Source data are provided as a Source Data file.

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