Fig. 7

Effects of TPX2 depletion on oocyte maturation, spindle assembly, chromosome alignment, and the expression of p-ADD1 and ADD1. A Immunoblot probed with anti-TPX2 antibody demonstrating the depletion efficiency of TPX2-specific morpholino (MO-TPX2) in mouse oocytes. B TPX2 protein content in control-MO and TPX2-MO-injected oocytes was quantified for three independent repeats. C Representative images of oocytes cultured in vitro for 16 h after treatment with control-MO or TPX2-MO. Bar, 100 μm. D Depletion of TPX2 caused meiotic cell cycle arrest in MI in mouse oocytes. E Subcellular localization of p-ADD1, spindle morphologies, and chromosome alignment after 16 h of maturation culture in oocytes injected with control-MO or TPX2-MO. Bar, 20 μm. F The rate of the oocyte with a normal spindle, aberrant spindle, or no spindle was quantified in the control-MO and ADD1-MO-injected oocytes. G The rate of the oocyte with normal p-ADD1 localization, abnormal p-ADD1 localization, or no p-ADD1 localization was recorded in the control and ADD1 disrupted oocytes. H The depletion of TPX2 downregulated the phosphorylation of ADD1 at S726 in mouse oocytes. I The protein level of p-ADD1 in control and TPX2-depleted oocytes was quantified for three independent repeats. J Effect of TPX2 depletion on ADD1 protein expression in mouse oocytes. (K) ADD1 protein levels in control-MO and TPX2-MO-injected oocytes were quantified for three independent replicates. In B, I, K, each bar denotes the mean ± SEM of the three independent repeats, and different uppercase letters or lowercase letters above columns indicate statistical difference at p < 0.01 or p < 0.05 by student’s t-test, respectively. 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