1. Long-term effects of repeated superovulation on ovarian structure and function in rhesus monkeys 
Guoying Dong, B.S., Yueshuai Guo, B.S., Huarong Cao, B.S., Tao Zhou, B.S., Zuomin Zhou, Ph.D., Jiahao Sha, Ph.D., Xuejiang Guo, Ph.D., Hui Zhu, Ph.D. 
Fertility and Sterility 
Volume 102, Issue 5, Pages e1 (November 2014)
DOI: /j.fertnstert
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Ovarian follicular morphologic analysis. (A) Hematoxylin and eosin staining reveals follicles at different developmental stages in each ovary, with no discernible abnormalities in cell morphology and histology. (: primordial follicle; : primary follicle; : secondary follicle; : antral follicle). Scale bar: 20 μm. (B) The proportion of follicles at each stage did not statistically significantly differ between the superovulation and normal groups (stage 0: primordial follicle; stage 1: primary follicle; stage 2: secondary follicle; stage 3: antral follicle; stage 4: atretic follicle). (C) Abnormal mitochondria were observed in the granulosa cells of the superovulation group, showing mitochondrial cristae degradation, mitochondrial matrix vague. Scale bar: 2 μm.
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3. Figure 2
Proliferation and apoptosis assessment of follicle cells. (A) Ki67-positive signals were mainly observed in the granulosa cells of secondary and antral follicles, and the ratio of Ki67-positive follicles did not statistically significantly differ between the superovulation and normal groups. Scale bar: 20 μm. (B) Terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick-end labeling (TUNEL)-positive signals were found mainly in granulosa cells. The ratio of TUNEL-positive cells did not statistically significantly differ between the two groups. Scale bar: 50 μm.
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4. Figure 3
Annotation of differentially expressed proteins. (A) Three differentially expressed proteins were selected for Western blot analysis; the trends in expression determined by Western blotting were very similar to those obtained by TMT quantification. S = superovulation group; N = normal group. ∗P<.05. (B) Subcellular distribution of differentially expressed proteins was based on gene ontology annotations published by the Database for Annotation, Visualization, and Integrated Discovery (DAVID). (C) Arrows indicate the functional annotations of proteins (red) and cell processes (yellow) as determined by Pathway Studio software.
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5. Supplemental Figure 1
Functional assessment of follicle cells. Estradiol (E2), progesterone (P4), and follicle-stimulating hormone (FSH) concentrations on (A) day 3 and (B) day 10 of the menstrual cycle. There were no statistically significant differences in these concentrations between the superovulation and normal groups. (C) The expressional changes of oocyte-specific genes in the ovarian tissues of superovulation monkeys compared with normal monkeys. The relative mRNA levels of five oocyte-specific genes showed no statistically significant differences between the superovulation and normal groups. (D) The expressional changes of estrogen synthesis-related molecules and enzymes in the ovarian tissues of superovulation monkeys compared with normal monkeys. The relative mRNA levels of StAR and CYP19A1 showed a statistically significant decrease in the superovulation group. ∗P<.05.
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions