Fig. 3

Effects of ADD1 deficiency on oocyte maturation, spindle assembly, and chromosome alignment. A Immunoblot probed with anti-ADD1 antibody demonstrating the depletion efficiency of ADD1-specific morpholino (MO-ADD1) in mouse oocytes. Protein loading was verified by the detection of β-actin. The molecular mass of target proteins is indicated on the right. B The ADD1 protein levels in control-MO- and ADD1-MO-injected oocytes were quantified for three independent repeats (normalized to β-actin, arbitrary units). In each set of experiments, the protein level of ADD1 was normalized to the value of oocytes in the control group. C Representative images of oocytes cultured in vitro for 16 h after control-MO or ADD1-MO treatment. Bar, 100 μm. D Depletion of ADD1 caused the failure of the first polar body extrusion in mouse oocytes. The rate of oocytes at the MI, AT1, and MII stages was quantified 16 h after meiotic resume in oocytes injected with control-MO and ADD1-MO, respectively. E Spindle morphologies and chromosome alignment 16 h after maturation culture in oocytes injected with control-MO or ADD1-MO. Green, α-tubulin; blue, DNA; Merge, overlapping of green and blue. Bar, 20 μm. F The rate of the oocyte with an aberrant spindle was recorded in the control-MO and ADD1-MO-injected oocytes. G The rate of the oocyte with misaligned chromosomes was quantified in the control and ADD1-depleted oocytes. Each column indicates the mean ± SEM of the three independent repeats in D, F, G. Different uppercase letters above columns denote statistical difference at p < 0.01 by student’s t-test in B, whereas bars that do not share the same uppercase letter are significantly different at p < 0.01 by the chi-square test in D, F, G