Mevastatin inhibits proliferation of rat ovarian theca–interstitial cells by blocking the mitogen-activated protein kinase pathway  Jakub Kwintkiewicz, Ph.D., Nastaran Foyouzi, M.D., Piotr Piotrowski, M.D., Ph.D., Izabela Rzepczynska, M.Sc., Antoni J. Duleba, M.D.  Fertility and Sterility  Volume 86, Issue 4, Pages 1053-1058 (October 2006) DOI: 10.1016/j.fertnstert.2006.04.020 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 (A) Effect of mevastatin in the absence and in the presence of serum on proliferation of theca–interstitial cells assessed by incorporation of radiolabeled thymidine; the cells were incubated for 48 hours without additives (control), with mevastatin (0.1, 0.3, 1, 3, and 10 μM), with 5% fetal bovine serum (FBS), or with a combination of mevastatin and FBS. (B) Effect of mevalonic acid on mevastatin-induced inhibition of proliferation of theca–interstitial cells assessed by incorporation of radiolabeled thymidine; the cells were incubated for 48 hours without additives (control), with mevalonic acid (10, 30, and 100 μM), with mevastatin (3 μM), or with a combination of mevalonic acid and mevastatin. Each bar represents mean ± SEM (N = 8). *Means significantly different from control (P<.01). †Means significantly different from FBS (P<.01). Kwintkiewicz. Proliferation of theca-interstitial cells. Fertil Steril 2006. Fertility and Sterility 2006 86, 1053-1058DOI: (10.1016/j.fertnstert.2006.04.020) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 (A) Proliferation of theca–interstitial cells in the absence and in the presence of ovarian steroids; the cells were incubated for 48 hours without additives (control), with progesterone and testosterone, all at 1 and 10 nM. (B) Effect of insulin and mevastatin on proliferation of theca–interstitial cells assessed by incorporation of radiolabeled thymidine; the cells were incubated for 48 hours without additives (control), with insulin (30 nM), with mevastatin (3 μM), or with combination of insulin and mevastatin. Each bar represents mean ± SEM (N = 8), means with no superscripts in common are significantly different. Kwintkiewicz. Proliferation of theca-interstitial cells. Fertil Steril 2006. Fertility and Sterility 2006 86, 1053-1058DOI: (10.1016/j.fertnstert.2006.04.020) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 (A) Effect of insulin and mevastatin on phosphorylation of ERK1/2 (pTpY 185/187) of theca–interstitial cells. (B) Effect of insulin and mevastatin on phosphorylation of Akt/PKB (pS 473) of theca–interstitial cells. The cells were incubated for 10 minutes without additives (control), with insulin (100 nM), with mevastatin (30 μM), or with a combination of insulin and mevastatin. Each bar represents mean ± SEM (N = 5). Means with no superscript letters in common are significantly different (P<.05). Kwintkiewicz. Proliferation of theca-interstitial cells. Fertil Steril 2006. Fertility and Sterility 2006 86, 1053-1058DOI: (10.1016/j.fertnstert.2006.04.020) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions