Total Protein and Heat Shock Proteins in FLF and BAL.

<p>Heat shock protein (HSP) Fetal Lung Fluid (FLF) Bronchoalveolar lavage fluid (BALF) Values: Mean ± SEM.</p><p>* p<0.05 vs PEEP and SI groups.</p><p>Total Protein and Heat Shock Proteins in FLF and BAL.</p

Pro-inflammatory cytokine and acute phase mRNA responses in the lung.

<p>(A) IL-1β did not increase with V_T (<i>p</i>>0.05). (B, C) IL-6 and IL-8 mRNA increased similarly for the V_T and V_T+SI groups, with no increase with SI relative to PEEP alone. (D, E) The acute phase genes Egr-1 and Cyr61 also increased with V_T and V_T+SI, with no increases with SI relative to PEEP alone. (F) CTGF mRNA did not change. *<i>p</i><0.05 vs PEEP only.</p

mRNA Fold increase over controls with 14 Day Ureaplasma exposure.

<p>* p&lt;0.05 vs saline controls.</p

In situ localization of EREG and AREG with LPS exposure and mechanical ventilation in lung.

<p>(<b>A–C</b>) EREG mRNA was not found in (A) unventilated controls or (B) IA LPS exposed lungs, but dramatically increased in peripheral lungs of lambs receiving (C) mechanical ventilation. EREG mRNA is localized to dome-shaped cells in peripheral lung (C, Insert). (<b>D–F</b>) AREG mRNA is found scattered in the peripheral lung of (D) Unventilated controls with a mild increase with (E) IA LPS exposure. (F) Mechanical ventilation increases the mRNA signal in similar cells to EREG. Scale bar = 50 µm</p

Groups, intra-amniotic (IA) exposures, gestational age and ventilation variables.

<p>(a) Gisslen <i>et al</i>. Innate immunity 2013 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096087#pone.0096087-Gisslen1" target="_blank">[12]</a>, (b) Collins <i>et al</i>. AJP-Lung 2010 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096087#pone.0096087-Collins1" target="_blank">[10]</a>, (c) Polglase et al. Pediatr Res 2010<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096087#pone.0096087-Polglase1" target="_blank">[11]</a></p

Angiotensin converting enzyme (ACE) 1 and 2 mRNA in lung with exposure to LPS, UP and mechanical ventilation.

<p>(<b>A–C</b>) mRNA values for unventilated controls (UVC), IA-LPS, and mechanical ventilation at 7 mL/kg (V_T7). (A) ACE1 was decreased by LPS but unaffected by V_T7. (B) ACE2 was increased by LPS and LPS+V_T7. (C) ACE2/ACE1 mRNA ratio increased with LPS and LPS+V_T7. * p&lt;0.05 vs UVC.</p

EGFR ligand mRNA response in lung to IA LPS, IA Ureaplasma, and mechanical ventilation.

<p>(<b>A–D</b>) mRNA response to IA-LPS and mechanical ventilation at 7 mL/kg (V_T7). (A) Amphiregulin (AREG) increases with LPS exposure with further increase with V_T7. (B) The Epiregulin (EREG) increase with V_T7 is not altered by LPS. (C) Heparin binding epithelial growth factor (HB-EGF) decreases with LPS exposure, but increases with V_T7. (D) Betacellulin (BTC) decreases with LPS and this decrease with LPS remains during V_T7. (<b>E–H</b>) mRNA response to IA-UP, UP+V_T7, mechanical ventilation at 15 mL/kg (V_T15) and V_T15+UP. (E) AREG and (F) EREG increase with V_T7 and V_T15. (G) HB-EGF increases with V_T7 and V_T15. (H) BTC had no change with UP or ventilation. * p&lt;0.05 vs unventilated, unexposed controls (UVC), † p&lt;0.05 vs UVC and IA-LPS, § p&lt;0.05 vs UVC and V_T7 without LPS</p

EGFR and Midkine mRNA fold changes.

<p>*p&lt;0.05 vs Controls; ^†p&lt;0.05 vs Controls and LPS alone.</p

Cytokine PCR Analysis.

<p>Quantitative PCR analysis of cytokines in fetal rhesus macaque skin following intra-amniotic exposure with LPS, IL-1β or UP and for different times of exposure (Panel a, b and c). Cytokine expression for LPS and LPS plus IL-1ra exposure groups (Panel d, e and f). Graphs represent mean mRNA fold change ± SD. <b>*</b>indicates a significant increase in transcript expression relative to untreated control group normalized to a value for 1.0. ɸ indicates a significant decrease in transcript expression relative to LPS at 16 hours.</p

Summary of PCR sets.

<p>Summary of PCR sets.</p