Fig. 4: TAp63α is synthesized in response to CDDP leading to the loss of primordial follicles.
From: Oocyte death is triggered by the stabilization of TAp63α dimers in response to cisplatin
A Immunoblot analysis of TAp63α and GAPDH at 2 and 6 h following treatment with CDDP, with or without pretreatment with cycloheximide (CHX). B Relative expression of TAp63α. Each dot represents a biologically distinct female ovary (n = 3 per group). C Histology of ovaries at 6 h post-exposure to CDDP or CDDP pretreated with CHX. D Immunofluorescence assay for TAp63α in ovaries. E Intensity analysis of TAp63α in oocytes of primordial follicles at 6 h (n = 3 mice per group). F Immunoblot analysis of TAp63α, PUMA, and GAPDH on SDS-PAGE, and TAp63α on BN-PAGE using ovarian extracts. An ovary from mice exposed to 0.45 Gy irradiation was used as a positive control for tetramer formation. 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 (a) Schematic of ovarian culture ex vivo. (b) Histological analysis of ovaries and immunofluorescence assay showing DDX4 and TAp63α. In H&E staining, intact primordial follicles and apoptotic ones are indicated by blue and green arrowheads, respectively. The middle and bottom images highlight the expression of DDX4 and TAp63α, respectively. Primordial follicles (PF) in immunofluorescence assay were marked by yellow arrowheads. H Total number of primordial follicles per ovaries after 96 h of culture (n = 8 for Ctr and CDDP groups; n = 12 for CHX + CDDP group). Statistical significance: n.s., not significant; *p < 0.05; ***p < 0.001; ****p < 0.0001.
