Mfn2 levels in two groups under the condition of normoxia and hypoxia.

<p><b>A and B</b>: The most representative image of western blotting for the protein expression level of Mfn2 and the relative signal intensity of Mfn2 protein levels in separate-culture group (trophoblasts) and co-culture group (MSC+trophoblasts). <b>C</b>: The comparison of Mfn2 mRNA levels in the two groups and the expression levels by the time in hypoxia. *<i>p</i><0.05 vs. the corresponding control group.</p

The apoptosis index were detected and the expression of apoptosis-related factors in two groups when incubated with hypoxia.

<p><b>A</b>: The most representative images of western blotting for Bcl-2, Bax, caspase 3 and caspase 9 in two groups; <b>B</b>: Comparison between Bcl-2 and Bax, caspase 3, caspase 9 protein levels; C: The most representative images of apoptosis in two groups; D The change of Apoptosis Index (AI) in two groups *<i>p</i><0.05 vs. the corresponding control group.</p

Expressions of hormone levels and function-related proteins of trophoblast cells in two groups.

<p><b>A</b>: The comparison ofβ-HCG levels of two groups in normoxia and hypoxia. <b>B</b>: The comparison of progesterone levels of two groups in normoxia and hypoxia. <b>C</b>: The most representative images of western blotting for MMP-2, MMP-9 and integrin β1 in two groups under the condition of hypoxia. *<i>p</i><0.05 vs. the corresponding control group.</p

The change of mitochondrial structure and function of two groups in normoxia and hypoxia.

<p><b>A</b>: The mitochondrial membrane potential (ΔΨm) was tested by using JC-1; <b>B</b>: Mitochondrial ultramicrostructrue were detected by TEM [Ba~Bd: the most representative image of mitochondrial morphology and cristae of trophoblast cells cultured alone or co-cultured in normoxia and hypoxia (black arrow); Ba’~Bd’: higher magnification.] <b>C</b>: ATP levels of the two groups in normoxia and hypoxia; *<i>p</i><0.05 vs. the corresponding control group.</p

Blastocyst formation rate.

<p>The blastocyst formation rate of the fertilized eggs collected from 5-week-old Kunming mice. The table presents the number of fertilized eggs collected from five mice. The values represent the rate of successful blastocyst development.</p><p>* <i>P</i>>0.05 <i>vs</i> control-siRNA transfected,</p><p>** <i>P</i><0.01 <i>vs</i> control-siRNA transfected.</p><p>Blastocyst formation rate.</p

Mfn2 expression in mouse fertilized eggs after transfection.

<p>Mfn2 mRNA expression was detected by qPCR and its level is significant lower in T group than that in C group (A). Mfn2 protein levels were determined by Western blotting (B). The quantitation of Mfn2 protein levels. C1 and T1: 4 cells fertilized eggs, C2 and T2: 8 cells fertilized eggs, C3 and T3: blastocyst. *<i>P</i><0.05. Data are means ± SD of 3 separate experiments (C).</p

Low Mfn2 expression attenuates the blastocyst formation rate and cleavage speed.

<p>In T group, the cleavage speed was reduced compared with that in control group, the blastocyst formation rate was significantly reduced in the T group compared with the C group. C1 and T1: 4-cell fertilized eggs, C2 and T2: 8-cell fertilized eggs. C3 and T3: Blastocyst. Data are means ± SD of 3 separate experiments.</p

Low <i>Mfn2</i> expression is associated with cell apoptosis in mice's ovarian tissues of POF.

<p>A: Apoptosis was determined by TUNEL in the ovarian tissues (light staining in the cytotrophoblasts of the control group; deeper staining in the cisplatin group); <b>B</b>: The most representative images of western blot for Bcl-2 and Bax; <b>C</b>: Comparison of the Apoptosis Index in the ovarian cells; <b>D</b>: Comparison between Bcl-2 and Bax protein levels; <b>E</b>: Comparison of Mfn2, Bcl-2, and Bax protein levels. *<i>P</i><0.05 vs control group.</p

Mfn2 distribution and Mfn2 levels in ovarian tissues of the two groups.

<p><b>A</b>: Mfn2 distribution was exhibited by immunohistochemistry and exclusively located in the cell's cytoplasm; <b>B</b>: The most representative image of western blotting; <b>C</b>: The relative amounts of Mfn2 protein levels in the control and cisplatin group; <b>D</b>: The comparison of Mfn2 mRNA levels in the ovarian tissue. *<i>P</i><0.05 vs control group.</p

Morphologic change in ovarian tissues and hormone levels of the two groups.

<p><b>A</b>: Morphology of ovarian tissues was determined by use of HE staining in both the control group and the cisplatin group (blue nuclei, red cytoplasm); <b>B</b>: Comparison of the ovarian weights; <b>C</b>: The comparison of hormone levels (FSH, Estradiol). *<i>P</i><0.05 vs control group.</p