Effect of LPA and COX inhibitors on the production of PGE2.

a<p>:p<0.001 vs the rest.</p><p>Uterine strips from rats pregnant on day 5 of gestation were incubated for 6 h with LPA 50 µM alone or in the presence of indomethacin 1 µM (a non selective COX-1 and COX-2 inhibitor) or NS-398 1 µM (a selective COX-2 inhibitor). In another set of experiments, the tissue was incubated for 6 h with indomethacin or NS-398 alone. The production of PGE2 was determined by radioimmunoassay.</p

Expression of LPA3 receptor and Lyso-PLD enzyme in the rat uterus during the window of implantation.

<p>LPA3 messenger (A) and protein (B) and Lyso-PLD messenger (D) and protein (E) were detected by RT-PCR and Western Blot. Also, LPA3 localization (C) was determined by immunhistochemistry in day 5. Black arrows indicate positive staining (200x). In A *** p<0.001 day 4 vs day 5, ** p<0,01 day 6IM vs day 6II. Results are shown as means ± sem. N = 4–6 for each point. d5: uterus from rats pregnant on day 5 of gestation; IM: implantation sites; II: inter-implantation sites.</p

Effect of LPA on the production of prostagandins in the rat uterus during the window of implantation.

<p>Uterine strips from rats pregnant on day 5 of gestation (implantation window) were incubated with LPA 50 µM for 6 h and the production of PGF2α (A) and PGE2 (B) was determined by radioimmunoassay. In B: *** p<0.001 vs C. Results are shown as means ± sem. N = 4–6 for each point. C: control.</p

Effect of LPA on decidualization and vascularization markers in the rat uterus during the window of implantation.

<p>Uterine strips from rats pregnant on day 5 of gestation (implantation window) were incubated with LPA 50 µM for 6 and 12 h and the expression of IGFBP-1 (A) and IL-10 (B and C) was studied. In A: *** p<0,001 vs the rest; In C: *** p<0,001 vs the rest, ** p<0.01 vs C. Results are shown as means ± sem. N = 4–6 for each point. C: control.</p

Effect of DGPP 10 µM on the production of PGE2 and on the expression of COX-2.

<p>Uterine strips from rats pregnant on day 5 of gestation were incubated for 6 h with DGPP 10 µM. The production of PGE2 was determined by radioimmunoassay. COX-2 mRNA and protein expression was analyzed by RT-PCR and Western Blot respectively.</p

RT-PCR primers and conditions.

<p>RT-PCR primers and conditions.</p

Effect of LPA on COX-1 and COX-2 expression in the rat uterus during the window of implantation.

<p>Uterine strips from rats pregnant on day 5 of gestation (implantation window) were incubated with LPA 50 µM for 6 h and the expression of COX-1 mRNA (A) and protein (B) and of COX-2 mRNA (C) and protein (D) was studied. In C: *** p<0,001 vs C; In D: ** p<0,01 vs C. Results are shown as means ± sem. N = 4–6 for each point. C: control.</p

First and the second antibody dilutions employed in Western Blot analyses.

<p>First and the second antibody dilutions employed in Western Blot analyses.</p

Effect of cannabinoid receptors’ antagonists on the production of PGE2 and on the expression of COX-2.

<p>Uterine strips from rats pregnant on day 5 of gestation were incubated for 6 h with AM251 10⁻⁸M (a selective CB1 antagonist) or AM630 10⁻⁸M (a selective CB2 antagonist). The production of PGE2 was determined by radioimmunoassay. COX-2 mRNA and protein expression was analyzed by RT-PCR and Western Blot respectively.</p

Endocannabinoids mediated LPA effect on the production of PGE2 and on COX-2 expression in the rat uterus during the window of implantation.

<p>Uterine strips from rats pregnant on day 5 of gestation (implantation window) were incubated with LPA 50 µM for 6 h in the presence of AM251 10<b>⁻</b>⁸M (a selective CB1 antagonist) or AM630 10<b>⁻</b>⁸M (a selective CB2 antagonist) or both. The production of PGE2 (A) and the expression of COX-2 mRNA (B) and protein (C) were determined. In A: *** p<0,001 vs C and 251+630; In B: *** p<0,001 vs the rest; In C: ** p<0,01 vs the rest. Results are shown as means ± sem. N = 4–6 for each point. C: control.</p