1. Concurrent estrogen action was essential for maximal progestin effect in oral contraceptives 
Yukiko Bono, M.D., Ph.D., Satoru Kyo, M.D., Ph.D., Tohru Kiyono, M.D., Ph.D., Yasunari Mizumoto, M.D., Ph.D., Mitsuhiro Nakamura, M.D., Ph.D., Yoshiko Maida, M.D., Ph.D., Masahiro Takakura, M.D., Ph.D., Hiroshi Fujiwara, M.D., Ph.D. 
Fertility and Sterility 
Volume 101, Issue 5, Pages (May 2014)
DOI: /j.fertnstert
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Effect of progestin and estrogen on the growth of immortalized epithelial cells from ovarian endometrioma. EMosis-CC/TERT1 or EMosis-CC/TERT1/ER cells were seeded into 6-well dishes and were treated for 72 hours with the factors contained in oral contraceptives (OCs). (A) 17β-ethynylestradiol (EE) at a concentration of 0.6 nmol/L, norethindorone (NET) at a concentration of 80 nmol/L, or both. (B) EE at a concentration of 0.6 nmol/L, levonorgestrel (LNG) at a concentration of 20 nmol/L, or both. (C) Dienogest at a concentration of 120 nmol/L, E2 at a concentration of 0.2 nmol/L, or both. Cell growth was monitored by counting cell numbers 72 hours after the treatment. Data are presented as mean ± SD of three independent experiments. *P<.05.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Western blot analysis of the expression of progesterone receptors (PRs). EMosis-CC/TERT1/ER or EMosis-CC/TERT1 cells were treated with various factors contained in oral contraceptives (OCs) for 72 hours; cell extracts were then recovered and subjected to Western blot analysis. EM-E6/E7/TERT/PRA or EM-E6/E7/TERT/PRB cells are immortalized endometrial epithelial cells in which PRA or PRB cDNA is stably expressed (23); these cells were used as a positive controls for PRA or PRB expression, respectively. Whereas EM-E6/E7/TERT/PRA cell extracts showed a clear PRA band on Western blots (94 kDa), EM-E6/E7/TERT/PRB extracts showed two bands when probed with an antibody (H-190) that recognizes both PRA and PRB; one band was of the expected size of intact PRB (114 kDa); the other band was located just below the PRA band (star), and was not a PRA band but a degraded PRB band; this supposition was confirmed by probing with a PRB-specific antibody (C1A2) in another Western blot analysis. (A) EMosis-CC/TERT1/ER cells lacked PRA or PRB expression in the absence of estrogen. The treatment with norethindorone (NET) at 80 nmol/L alone caused slight induction of both PRA and PRB. However, coadministration of 17β-ethynylestradiol (EE; 0.6 nmol/L) and NET resulted in significant induction of both PRA (94 kDa) and PRB (114 kDa). A similar combinatorial effect was observed in cells treated with LNG (20 nmol/L) and EE (0.6 nmol/L), although levonorgestrel (LNG) alone at 20 nmol/L had no effect on expression of either PR. (B) EMosis-CC/TERT1 did not exhibit any induction of PR expression even with combinatorial treatments.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Immunohistochemical analyses of progesterone receptor (PR) A, PRB, and estrogen receptor (ER) α expression in ovarian endometrioma. (A) The surgical specimens of ovarian endometriomas from ten patients were immunohistochemically analyzed for PRA, PRB, and ERα expression. The percentages of epithelial cells with nuclear staining among all epithelial cells throughout the section were scored as follows: 1 (<10%), 2 (10%–24%), 3 (25%–49%), and 4 (≥50%). (B) Representative staining patterns are shown; these examples are each from case 7. HE = hematoxylin and eosin. (C) Analysis of variance of PRA and PRB for ERα. PRB expression and ERα expression were significantly correlated (r = 0.046).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions