Fig 1: Cpeb3 mutation causes POI in mice by downregulation of the oocyte-derived factor, Gdf9.A Western blot analysis of GAPDH, GDF9, PCNA, and Cleaved-Caspase3 protein expression in 3-month-old WT and Cpeb3-mutant ovaries. GAPDH was used as the internal control. B Western blot analysis of GDF9 and GAPDH in WT and Cpeb3-mutant oocytes. GAPDH was used as the internal control. C Ovarian sections stained for GDF9 at 3-month-old WT and Cpeb3-mutant ovaries. D Gdf9 mRNA levels were performed by qRT-PCR in WT and Cpeb3-mutant oocytes. Gapdh was used as the internal control. Scale bar = 50 μm and 25 μm in C. ***P < 0.001.
Fig 2: Immunohistochemistry of the chimeric ovaries from NANOS3−/− cloned embryos.The histology of chimeric (#1, a–d) and nonchimeric (#2, e–h) fetal ovaries by ESR1 (a,e), NANOS3 (b,f), GDF9 (c,g) and VASA (d,h) immunostaining was compared. Arrowheads indicate primordial follicles (a–d). Although it appeared to be single layer of follicular epithelium-like cells, no germ cells were observed in foetus #2 ovary in (e–h). Scale bars in (a–c,e–g) = 50 μm; and in (d,h) = 25 μm.
Fig 3: The transcriptional dynamics of DEHP decaying primordial follicle formation. The middle line marks the time window of primordial follicle formation, which represents germ cell fate transition from germline cysts to follicles (right annotation text), however, it consists of three main biological events, granulosa cell invasion, cyst breakdown, and follicle assembly (left annotation text). The left panel indicates normal follicle formation without DEHP, consisting of normal expression levels of Figla, Lhx8, and Zp3, and TGF-beta members (Gdf9, Bmpr1a, and Smad4) in germ cells, as well as TGF-beta receptors (Bmpr1a) and effectors (Smad3 and Smad4) in pre-granulosa cells. The right panel displays follicle assembly impaired by DEHP, which shows the decreasing transcripts of key genes for follicle formation (Figla, Lhx8, and Zp3) and of TGF-beta components in germ cells (Gdf9) and pre-granulosa cells (Bmpr1a and Smad3), but the increasing levels of Smad4 in both cell types and Bmpr1a only in germ cells.
Fig 4: Co-treatment of KP and LiCl during IVM and their effects on oocyte competence. (A) Expression levels of GDF9 and BMP15 in mature porcine oocytes. More than 20 oocytes from each group were used for the staining. X400 Magnification. (B) Treatment with KP significantly increased the competence of the oocytes, while treatment with LiCl decreased the competence. (C) Co-treatment of KP and LiCl. KP and LiCl had significant inverse effects on the cleavage rate and blastocyst formation rate compared to those observed in the control group, and the KP-treated group had the highest total number of blastocysts among all groups. A total of six replicates were performed. Bars with letters indicate significant differences (P < 0.05).
Fig 5: The protein levels were analyzed by immunocytochemistry in porcine oocytes. Expression of (A) GDF9; (B) BMP15; (C) SOD1; (D) CDK1; (E) PGC1α in oocytes, respectively. Within the same protein, bars with an asterisk are significantly different (p < 0.05). Results are shown as the average ± SEM of at least three repeats of independent experiments. Ramelteon concentration is 10−11 M. Scale bars = 40 μM.
Supplier Page from Abcam for Anti-GDF 9 antibody