1. mTOR kinase inhibition results in oocyte loss characterized by empty follicles in human ovarian cortical strips cultured in vitro 
Marie McLaughlin, Ph.D., Pasquale Patrizio, M.D., Umit Kayisli, Ph.D., Janelle Luk, M.D., Travis C. Thomson, Ph.D., Richard A. Anderson, M.D., Ph.D., Evelyn E. Telfer, Ph.D., Joshua Johnson, Ph.D. 
Fertility and Sterility 
Volume 96, Issue 5, Pages e1 (November 2011)
DOI: /j.fertnstert
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Human oocyte loss results from mTOR inhibition without an induction of apoptosis. (A) Plot of raw numbers of follicles found in cortical strips that were either fixed immediately upon isolation from patient or cultured for 6 days in the presence of ethanol vehicle (VEH) or 100 nM mTOR inhibitor rapamycin (RAP). (B) Representative section from intact follicle from RAP-treated cortical strip. (C) Example of “shrunken” oocyte phenotype; compare to oocyte in B. Bar = 50 μm. (D) Example of “empty” follicle. Representative central section from follicle lacking oocyte from RAP-treated cortical strip. Bar = 50 μm.
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
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3. Figure 2
Caspase activity measurements from whole ovarian strips 1, 3, and 6 days (1d, 3d, 6d) after treatment. Compared to freshly (FRESH) isolated cortical strips (left-most bars), rapamycin (RAP) treatment did not result in a significant increase in the activity of caspases 3/7 (A), 8 (B), or 9 (C). Camptothecin (CT; 4 μM, right-most bars on each graph) was used as a positive control for caspase activation, and induced significantly increased caspase activity in each case (significance denoted by lowercase letters, P<.00005, analysis of variance (ANOVA) followed by Tukey’s HSD test).
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
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4. Supplemental Figure 1
Hypothetical scheme for the generation of empty follicles. Granulosa cells (GC) are known to differentiate into the mural and cumulus lineages. A third lineage, phagocytic type of GCs, is proposed, termed phagocytic GCs, which normally serve to survive and clear the oocyte and dead and dying GCs during classic follicle atresia. However, if phagocytic GCs arise and phagocytose the oocyte in the absence of concurrent apoptosis, an empty follicle would be produced. Although phagocytic GCs can destroy intact, “healthy” oocytes in fruit flies, this question has not been addressed in mammals.
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions