1. Effect of hormonal agents on monocyte chemotactic protein-1 expression by endometrial epithelial cells of women with endometriosis 
Annie Boucher, M.S., André Lemay, M.D., Ph.D., Ali Akoum, Ph.D. 
Fertility and Sterility 
Volume 74, Issue 5, Pages (November 2000)
DOI: /S (00)

2. Figure 1
Dose-dependent course (A and B) and time course (C and D) of monocyte chemotactic protein (MCP)-1 expression by IL-1β–treated endometrial epithelial cells. Confluent cultures were exposed to different concentrations of IL-1β (0.01–10 ng/mL) for increasing periods of time. At each time point, the culture supernatant was recovered for measurement of MCP-1 protein secretion by ELISA, and total cellular RNA was extracted to evaluate MCP-1 mRNA expression by using Northern blot analysis. (A) and (C) show MCP-1 protein secretion (expressed in pg/mL). (B) and (D) are autoradiographs showing hybridization of MCP-1 messenger RNA with 32P-labeled MCP-1 complementary DNA probe. Hybridization of 28S ribosomal RNA with a 32P-labeled 28S complementary DNA probe showed equal RNA loading (representative data from patient 1, Table 1).
Boucher. MCP-1 regulation by hormonal agents. Fertil Steril 2000.
Fertility and Sterility  , DOI: ( /S (00) )

3. Figure 2
Effect of danazol on interleukin (IL)-1β–induced monocyte chemotactic protein (MCP)-1 expression by endometrial epithelial cells. Cells were treated with different concentrations of danazol (10−7 M–10−5 M) (see Materials and Methods) and with or without IL-1β, 0.1 ng/mL, for 6 hours. (A), Secretion of MCP-1 protein in the culture medium was determined by using ELISA and is expressed as a percentage of control (conditioned medium from cultures were not subjected to hormonal treatment). Values are means (±SE) obtained from duplicate determinations in culture from four different endometrial biopsy specimens (patients 4, 5, 6, and 7, Table 1). ∗∗Significantly different from control (P<.01). (B), MCP-1 messenger RNA steady-state levels in endometrial cells were analyzed by using Northern blot analysis; an autoradiograph from a representative experiment is shown (patient 4, Table 1). Concomitant hybridization of 28S ribosomal RNA demonstrated equal RNA loading.
Boucher. MCP-1 regulation by hormonal agents. Fertil Steril 2000.
Fertility and Sterility  , DOI: ( /S (00) )

4. Figure 3
Effect of dexamethasone on monocyte chemotactic protein (MCP)-1 expression by endometrial epithelial cells. Cells grown to confluence were incubated with different concentrations of dexamethasone (DXS) (10−12M–10−6M) for 18 hours and then with interleukin (IL)-1β, 0.1 ng/mL, for 6 more hours. Conditioned media and cells were then recovered for ELISA and Northern blot analyses of MCP-1 protein and messenger RNA expression, respectively. (A), Secretion of MCP-1 protein was expressed as a percentage of control (cultures stimulated with IL-1β, 0.1 ng/mL, without previous incubation with DXS). Values are means (±SE) obtained from duplicate determinations in cultures from three different endometrial biopsy specimens (patients 8, 9, and 10, Table 1). ∗P<.05 compared with control; ∗∗P<.01 compared with control. (B), Representative autoradiograph showing MCP-1 messenger RNA synthesis (patient 8, Table 1) and 28S ribosomal RNA showing equal RNA loading.
Boucher. MCP-1 regulation by hormonal agents. Fertil Steril 2000.
Fertility and Sterility  , DOI: ( /S (00) )

5. Figure 4
Effect of a GnRH agonist (Suprefact) on monocyte chemotactic protein (MCP)-1 expression by endometrial cells. Cells were treated with agonist vehicle (benzalkonium chloride) (seeMaterials and Methods) and then with or without interleukin (IL)-1β for 6 hours. Secretion of MCP-1 protein in the culture medium was determined by using ELISA and is expressed as a percentage of control (conditioned medium from cultures were not subjected to hormonal treatment). (A), Values are means (±SE) obtained from duplicate determinations in cultures from three different endometrial biopsy specimens (patients 11, 12, and 13, Table 1). (B), MCP-1 messenger RNA steady-state levels in endometrial cells were analyzed by using Northern blot analysis; an autoradiograph of a representative experiment is shown (patient 16, Table 1). Concomitant hybridization of 28S ribosomal RNA demonstrated equal RNA loading.
Boucher. MCP-1 regulation by hormonal agents. Fertil Steril 2000.
Fertility and Sterility  , DOI: ( /S (00) )