1. Selective blockade of prostaglandin E2 receptors EP2 and EP4 signaling inhibits proliferation of human endometriotic epithelial cells and stromal cells through distinct cell cycle arrest 
JeHoon Lee, M.S., Sakhila K. Banu, Ph.D., Royce Rodriguez, B.S., Anna Starzinski-Powitz, Ph.D., Joe A. Arosh, Ph.D. 
Fertility and Sterility 
Volume 93, Issue 8, Pages (May 2010)
DOI: /j.fertnstert
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Effects of inhibition of EP1, EP2 and EP4 receptors on proliferation of human endometriotic epithelial and stromal cells: dose response experiment for EP1, EP2 and EP4 inhibitors in endometriotic epithelial cells 12Z (A) and endometriotic stromal cells 22B (B). Pharmacological inhibition of EP2, EP4 or EP2/EP4 decreased proliferation of 12Z cells (C) and 22B cells (D), ∗Control (CONT) vs. EP2-I, EP4-I or EP2-I/EP4-I; P<0.05. Pharmacological inhibition of EP2, EP4 or EP2/EP4 decreased viability of 12Z cells (E) and 22B cells (F), ∗Control (CONT) vs. EP2-I, EP4-I or EP2-I/EP4-I; P<0.05. Knock-down of EP2, EP4, EP2/EP4 genes decreased proliferation of 12Z cells (E) and 22B cells (F), ∗Control (CONT) vs. EP2-, EP or EP2-/EP4-; P< Numerical values are expressed as the mean ± SEM of three independent experiments. See “Materials and Methods” for more details.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Effects of inhibition of EP2/EP4 on regulation of cell cycle in human endometriotic epithelial and stromal cells. Inhibition of EP2/EP4 arrests progression of endometriotic epithelial cells 12Z through G1–S and G1–M checkpoints (A) and endometriotic stromal cells 22B through G2–M checkpoint (B). Control versus EP2-I/EP4-I, ∗P<.05. Western blot analyses on expression and regulation of CDK4, CDK6, CDK2, and CDK1 (C–F), cyclin D1, D2, D3, E2, A, and B1 (G–J), and CDK inhibitor p15, p16, p21, and p27 (K–N) proteins in 12Z and 22B cells. Control versus EP2-I/EP4-I, ∗P<.05. Numerical values are expressed as the mean ± SEM of three independent experiments. Please see Materials and Methods section for more details. CONT = control.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Selective inhibition EP2 and EP4 decreases proliferation of human endometriotic cells but not of eutopic endometrial cells. (A) Ectopic endometriotic epithelial cells12Z. a: control versus PGE2; b: control versus EP2/EP4-I; c: PGE2 versus PGE2 + EP2/EP4-I; P<.05. (B) Eutopic endometrial epithelial cells (HES) from endometriosis-free women. (C) Ectopic endometriotic stromal cells 22B. d: control versus PGE2; e: control versus EP2/EP4-I; f: PGE2 versus PGE2 + EP2/EP4-I; P<.05. (D) Eutopic endometrial stromal cells (HESC) from endometriosis-free women. (E) PGE2 production by 12Z, HES, 22B, HESC at basal condition. g: 12Z versus HES; h: 22B versus HESC; P<.05. Numerical values are expressed as the mean ± SEM of three independent experiments. Please see Materials and Methods section for more details.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Proposed EP2/EP4-mediated PGE2 signaling on regulation of cell cycle in human endometriotic epithelial and stromal cells. (A) Inhibition of EP2/EP4 decreased proliferation of human endometriotic epithelial cells 12Z through [1] down-regulation of G1-specific cyclins D1, D2, D3, and CDK4 and CDK6; [2] down-regulation of G1–S transition-specific cyclin E2 and CDK2; [3] down-regulation of S-specific cyclin A and CDK2; [4] down-regulation of G2-specific cyclin A and CDK1; and [5] down-regulation of M-specific cyclin B1 and CDK1. Deregulated assembly between cell-specific CDKs and their cyclins (1 and 2) inhibits progression of cells through G1 and from G to S phases and thus leads to cell cycle arrest at G1–S checkpoint (6). Deregulated assembly between cell-specific CDKs and their cyclins (3, 4, and 5) inhibits progression of cells through G2 and M and results in cell cycle arrest at G2–M checkpoint (7). (B) Inhibition of EP2/EP4 decreased proliferation of human endometriotic stromal cells 22B through [1] down-regulation of S-specific cyclin A and CDK2; [2] down-regulation of G2-specific cyclin A and CDK1; and [3] down-regulation of M-specific cyclin B1 and CDK1. Deregulated assembly between cell-specific CDKs and their cyclins (1, 2, and 3) inhibits progression of cells through G2 and M and results in cell cycle arrest at G2–M checkpoint (4).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions