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Fig. 1 | Reproductive Biology and Endocrinology

Fig. 1

From: Insufficient HtrA2 causes meiotic defects in aging germinal vesicle oocytes

Fig. 1

Reduced expression of HtrA2 in aged oocytes. A Representative western blot showing HtrA2 expression in oocytes from young and old mice. The relative amount of HtrA2 in old mice was estimated based on the level of actin, which was used as control. Statistical analysis of western blotting in GV oocytes from aged or young mice. Normalized band intensities: HtrA2/Actin. HtrA2 protein levels were significantly lower in oocytes obtained from old mice compared to young mice. Data are expressed as the mean ± SEM of three independent experiments. *, p < 0.05. B Diagram summarizing the HtrA2-KD approach using siRNA microinjection. C Western blotting verification of endogenous HtrA2 knockdown using two different siRNAs and 100 fully grown GV-stage oocytes per lane. The relative amount of HtrA2 in HtrA2-KD oocytes was estimated based on the level of actin and/or glyceraldehyde 3-phosphate dehydrogenase (GAPDH). D Phase-contrast images of control and HtrA2 siRNA injected oocytes. Yellow arrowheads point to oocytes with symmetrical division or a large polar body. Red arrows point to oocytes with no polar body. Scale bar: 80 μm. E–G Quantitative analysis of the GVBD rate, extrusion frequency of the first polar body (Pb1; control n = 341; siRNA-1 n = 255; siRNA-2 n = 298), and percentage of oocytes arrested at metaphase I (control n = 164; siRNA-1 n = 167; siRNA-2 n = 153) in control and HtrA2-KD oocytes. Data are expressed as the mean ± SEM from three independent experiments. *, p < 0.05. ns, no significant difference

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