1. Uterine luminal epithelium–specific proline-rich acidic protein 1 (PRAP1) as a marker for successful embryo implantation 
Honglu Diao, Ph.D., Shuo Xiao, M.S., Fei Zhao, B.S., Xiaoqin Ye, M.D., Ph.D. 
Fertility and Sterility 
Volume 94, Issue 7, Pages e1 (December 2010)
DOI: /j.fertnstert
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2. Figure 1
(A) Expression levels of Prap1 in wild-type (WT; +/+) and Lpar3−/− (−/−) peri-implantation uterus using real-time polymerase chain reaction (PCR). The inset figure shows the relative Prap1 expression levels in days 3.5 and 4.0 WT and Lpar3−/− uterus. D: days; IS: implantation site; NIS: non-implantation site. (B) Localization of Prap1 mRNA in days (D) 0.5, 3.5, 4.5, and 5.5 WT (+/+) and days 4.5 and 5.5 Lpar3−/− (−/−) uterus by in situ hybridization using a Prap1 antisense probe. No specific signals were detected using a sense probe (data not shown). (C) Time course effect of progesterone (P) on Prap1 expression in the uterus of WT ovariectomized mice determined by real-time PCR. (D) Time course effects of 17β-estradiol (E2) alone or cotreatment with P (treatment time indicated as hours) on Prap1 expression in the uterus of WT ovariectomized mice determined by real-time PCR. Data were first normalized by the relative expression of loading control glyceraldehyde 3-phosphate dehydrogenase (GAPDH), then compared with the oil-treated control at respective time points and expressed as fold change relative to control. Similar results were obtained when a different house-keeping gene, hypoxanthine phosphoribosyltransferase 1, was used as a loading control (data not shown). (E) Expression of Prap1 mRNA in WT uterus with delayed and activated embryo implantation by in situ hybridization using a Prap1 antisense probe. (F) Expression of Prap1 mRNA in WT uterus with artificial decidualization by in situ hybridization using a Prap1 antisense probe. A, C, D: ∗P<.05 (unequal variance two-tail Student t test); n = 3–6; error bar = SD. B, E, F: Sections were counterstained with 1% methyl green (9, 10); red asterisk = embryo; Dec = decidual zone; LE = luminal epithelium; Myo = myometrium; S= stroma; scale bars: 100 μm (B, E), 200 μm (F).
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
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3. (A) Quantification of Prap1 expression levels in day 4
(A) Quantification of Prap1 expression levels in day 4.5 WT (+/+) and Lpar3−/− (−/−) uterine luminal epithelium (LE) and whole uterus using real-time polymerase chain reaction (PCR). (B) Up-regulation of Prap1 mRNA in WT ovariectomized uterus treated with E2 for 3 days by in situ hybridization using a Prap1 antisense probe. Scale bar: 100 μm. (C) Undetectable Prap1 mRNA in day 3.5 and day 4.5 pseudopregnant WT uterus by in situ hybridization using a Prap1 antisense probe. Scale bar: 200 μm. (B, C) Sections were counterstained with 1% methyl green (9, 10). (D) Quantification of Prap1 expression levels in experimentally delayed- and activated-implantation WT uterine luminal epithelium (LE) and whole uterus using real-time PCR. (A, D) ∗P<.05 (unequal variance two-tail Student t test); n = 6; error bar = SD.
Fertility and Sterility  , e1DOI: ( /j.fertnstert )
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