Maternal IL-6 plasma levels and spleen weight in mice exposed to chronic LPS treatments.

<p>Chronic LPS (5 µg/Kg) treatment was performed daily from E11.5 until E15.5. (A) Maternal plasma was extracted 4 h after last LPS treatment on E15.5 (Vehicle n = 9; LPS n = 10). (B) Maternal splenic weight from animals exposed to chronic LPS treatment (Vehicle n = 9; LPS n = 10). Values are means±SEM. *P<0.05, **<i>p<</i>0.001 (one-way ANOVA followed by the Tukey’s post-hoc test).</p

Placental and maternal myocardial P-gp activity after acute sub-lethal LPS exposure:

<p>Fetal Units [³H]digoxin accumulation (4 fetal units/dam were randomly harvested and assayed) on E15.5 (A) (n = 5 dams/gp) and E17.5 (B) (4 h n = 5 dams/gp; 24 h n = 4 dams/gp); 4 or 24 h after acute LPS treatment. Maternal myocardial [³H]digoxin accumulation on E15.5 (C) and E17.5 (D) 4 or 24 h after acute LPS treatment. Values are means±SEM. *<i>p<</i>0.05 (one-way ANOVA followed by the Tukey’s post-hoc test on E15.5 and Kruskal-Wallis analysis of variance followed by Dunn’s test on E17.5).</p

Rate of fetal death/reabsorption after acute LPS exposure.

<p>Rate of fetal death/reabsorption after acute LPS exposure.</p

Placental size and P-gp transport efficiency.

<p>Small, mid-range and larger placentas from each litter were grouped and averaged for placental (A) and fetal (B) weight and fetal to placental (F:P) weight ratio (C). (D) shows the relationship between individual placental weights and [³H]digoxin fetal accumulation at E15.5, for vehicle (r = −0.6657, n = 31 fetuses from 5 litters, <i>p</i><0.0001) and chronic LPS (r = −0.5099, n = 39 fetuses from 6 litters, <i>p</i><0.001) groups. (E) [³H]digoxin fetal accumulation from small, mid-range and larger placentas at E15.5 (vehicle n = 5; LPS n = 6). (A,B,C and E, repeated measures two-way ANOVA followed by Bonferonni’s test, <i>p</i><0.05; Pearson’s correlation test).</p

Placental mRNA expression of the multidrug resistance genes (Abcb1a, Abcb1b, and Abcg2), Il-6, Tnf-α, Il-10 and Tlr-4 after chronic LPS exposure.

<p>The small, mid-range and larger placentas from each litter were grouped and assayed for mRNA expression. (Veh, n = 7 dams; LPS n = 7). (Repeated measures two-way ANOVA followed by Bonferonni’s test, <i>p</i><0.05). Relative gene expression normalized to <i>Tbp, Gapdh</i> and <i>Hprt</i>.</p

Rate of of fetal death/reabsorption after multiple LPS exposure.

*<p>Dams received daily Veh/LPS treatments from E11.5 until E15.5 and were euthanized 4 hs after last treatment on E15.5.</p

Placental mRNA expression of the multidrug resistance genes (Abcb1a, Abcb1b, and Abcg2) and the pro-inflammatory cytokine Il-6 after acute sub-lethal LPS exposure: one placenta per group was randomly harvested 4 h after LPS insult and processed for qPCR analyses (n = 6 dams/gp).

<p>Values are fold increase of the means±SEM. *<i>p<</i>0.05 (one-way ANOVA followed by the Tukey’s post-hoc test). Relative gene expression normalized to Tbp Gapdh.</p

Maternal IL-6 plasma levels in mice exposed to acute LPS treatment.

<p>Vehicle and acute LPS (150 µg/Kg) treatments were performed and maternal plasma was extracted: 4 h (n = 6/gp) or 24 h (n = 5/gp) after single LPS exposure. Values are means±SEM. ***<i>p<</i>0.0001 (one-way ANOVA followed by the Tukey’s post-hoc test).</p

Placental and maternal myocardial P-gp activity after chronic sub-lethal LPS exposure.

<p>(A) fetal units [³H]digoxin accumulation (all fetuses/dam) and (B) maternal myocardial [³H]digoxin accumulation, 4 h after last LPS chronic treatment (daily from E11.5 until E15.5) on E15.5 (vehicle n = 5; chronic LPS n = 6). Values are means±SEM. *P = 0.05 (one-way ANOVA followed by the Tukey’s post-hoc test).</p

Distribution of forces on the zona pellucida, blastomeres or external environment.

<p>Individual blastomeres, especially those located most inferiorly, experience a significant force on the order of 1 nN. The extreme stiffness of the ZP relative to the weight of the embryo results only in minimal modification of the zona, when cultured in polystyrene. It will be the interface between the ZP and the external surface that will deform differently according to the characteristics of the external surface (F_Z-ex). This is because only the softer of two materials will yield to pressure <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041717#pone.0041717-Popov1" target="_blank">[45]</a> (<b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041717#pone.0041717.s003" target="_blank">Figure S3</a></b>). For example, the weight of the embryo on polystyrene will indent the ZP of 20–120 nm (0.1% to 0.7%) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041717#pone.0041717-Murayama1" target="_blank"></a><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041717#pone.0041717-Bertrand1" target="_blank">[34,46]</a>. In contrast, when embryos are cultured on collagen gel, the collagen itself will yield (up to 1 µm) to the weight of the embryo and the ZP will not indent. Indentation is defined as the linear deformation experienced in the radial axis either by the embryo’s zona pellucida or by the substrate (collagen or PDMS). Sed: sedimentation time.</p>†<p> <i>From </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041717#pone.0041717-Murayama1" target="_blank">[<i>34</i>]</a><i>.</i></p