Fig. 4

Premature upregulation of TAp63 in GSK-3β-inhibited ovary. a The expression pattern of γ-H2AX in fetal and neonatal mouse ovary in vivo. Mouse ovaries from 13.5 dpc, 15.5 dpc, 17.5 dpc, and 1 dpp were immunostained for γ-H2AX (green) and DDX4 (red). The nucleus was stained by Hoechst (blue). γ-H2AX displayed intensive expression in the germ cell nucleus from 15.5 to 17.5 dpc. b The expression pattern of TAp63 in fetal and neonatal mouse ovary in vivo. Mouse ovaries from 13.5 dpc, 15.5 dpc, 17.5 dpc, 18.5 dpc, and 1 dpp were immunostained for TAp63 (green) and DDX4 (red). The nucleus was stained by Hoechst (blue). The TAp63 protein located within somatic cells in fetal ovary and began to express in the germ cell nucleus from 18.5 dpc afterward. c–e TAp63 expression was upregulated in fetal ovary and displayed premature localization within the oocyte nucleus following GSK-3β inhibition. Before the examination, ovaries of 14.5 dpc were cultured in vitro with DMSO or BIO for 3 days. c qRT-PCR analysis of TAp63 mRNA level following GSK-3β inhibition (normalized to β-actin) (Additional file 8: Individual data values). d Western blotting analysis of TAp63 protein level following GSK-3β inhibition. GAPDH was used as an internal control. e Immunofluorescence staining for TAp63 (green) and DDX4 (red). The nucleus was stained by Hoechst (blue). TAp63 showed premature expression within the oocyte nucleus following GSK-3β inhibition (arrowhead). f qRT-PCR results showed that relative mRNA expression level of p21, Bad, and Noxa increased significantly in GSK-3β-inhibiting ovaries (Additional file 8: Individual data values). The data are presented as mean ± s.d. The asterisk (*) denotes a statistically significant difference between the control and treatment groups. *P < 0.05, **P < 0.01, and ***P < 0.001 (t test). Scale bars, 200 μm