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Macrophage Scavenger Receptor(s) and Oxidatively Modified Proteins in Endometriosis
Ana A. Murphy, M.D., Wulf Palinski, M.D., Sara Rankin, Ph.D., Arlene J. Morales, M.D., Sampath Parthasarathy, Ph.D. Fertility and Sterility Volume 69, Issue 6, Pages (June 1998) DOI: /S (98)
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FIGURE 1 Degradation of LDL by peritoneal cells. Cells obtained from the PF of 13 women with endometriosis and 10 control patients were plated at 3 × 106 cells/well in six-well plates. After overnight culture in RPMI medium with 10% fetal bovine serum, cells were washed and incubated with 5 μg 125I-labeled of native LDL, EC-LDL, or Cu-LDL. The medium was assayed for noniodide, trichloroacetic acid soluble radioactivity. Values are expressed as micrograms of lipoprotein degraded in 5 hours per milligram of macrophage cell protein. ■ = endometriosis; □ = controls. Fertility and Sterility , DOI: ( /S (98) )
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FIGURE 2 Incorporation of 14C-oleate into cholesterol esters by peritoneal cells. Peritoneal cells of women (n = 5) with endometriosis incorporated significantly more oleate when incubated with EC-LDL (P <0.002) or Cu-LDL (P <0.02) than with native LDL. Values are expressed as nanomole of oleate incorporated per milligram of cell protein. Fertility and Sterility , DOI: ( /S (98) )
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FIGURE 3 Western blot of PF from women with endometriosis and from controls. Peritoneal fluids were separated on a 4%–20% SDS polyacrylamide gel, transblotted to a nitrocellulose membrane, and stained with MDA2, the monoclonal antibody to MDA-lysine (A) and HNE-7, the antiserum against 4-HNE-lysine (B), as described in Methods. Lanes 1 and 2: PF from control patients. Lanes 3–6: PF from women with endometriosis. Both antibodies recognize oxidation-specific epitopes on several protein bands. The amount of protein applied to each lane is identical (10 mg), and the Western blot therefore does not reflect quantitative differences between control patients and patients with endometriosis. Fertility and Sterility , DOI: ( /S (98) )
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FIGURE 4 Peritoneal fluid from women with endometriosis competes with labeled Ox-LDL for degradation by mouse peritoneal macrophages. Peritoneal fluid, added to macrophage cultures containing labeled copper oxidized LDL (Cu-LDL) or endothelial cell modified LDL (EC-LDL), shows a dose-dependent decrease in macrophage degradation of labeled LDL. ■ = EC-LDL and □ = Cu-LDL. Fertility and Sterility , DOI: ( /S (98) )
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