Fig. 3: CDDP induces the accumulation of TAp63α in oocytes of primordial follicles ex vivo without tetramer formation.
From: Oocyte death is triggered by the stabilization of TAp63α dimers in response to cisplatin
A Histology and DAB staining for Dead-box helicase 4 (DDX4), an oocyte marker [26], in ovaries treated with solvent (Control, Ctr) or 20 μM CDDP for 24 h. Intact primordial follicles and apoptotic ones are indicated by blue and green arrowheads, respectively. B Expression of γH2AX, a DNA damage marker, and cPARP, an apoptotic marker, in ovaries cultured with either solvent (Control, Ctr) or 20 μM CDDP for 24 h. Pink arrowheads indicate the signals. C Total numbers of primordial follicles per ovaries (n = 4). D Immunoblot analysis showing time-dependent expression of TAp63α and GAPDH. E Quantification of TAp63α intensity, expressed as fold changes. Each dot represents a biologically distinct female ovary (n = 3 in each group). F Immunoblot analysis for TAp63α on BN-PAGE using ovaries treated with solvent (Control, Ctr) and 20 μM CDDP, harvested at 6 and 16 h. An ovary exposed to 0.45 Gy irradiation for 4 h was used as a positive control. The molecular weights of the TAp63α dimer (480 kDa, a black arrowhead) and tetramer (720 kDa, a red arrowhead) are indicated on the BN-PAGE. G Histology and immunofluorescence assay of TAp63α in ovaries cultured with solvent (Control, Ctr) or 20 μM CDDP for 6 h. Insets show TAp63α expression in oocytes of primordial follicles. H Intensity analysis of TAp63α in primordial follicles (n = 30) at 6 h post-CDDP treatment. I Immunoblot analysis of TAp63α and GAPDH at each time point. Phosphorylated and unphosphorylated bands on SDS-PAGE are marked with red and black arrowheads, respectively. Statistical significance: n.s. not significant; *p < 0.05; ****p < 0.0001.
