Expression of Snail1 in tumor and stroma according to tumor stage.

<p>Snail1 immunoreactivity was determined in the stromal or carcinoma cells corresponding to colorectal tumours classified in the different stages. According to Snail1 expression, tumours were classified as presenting Snail1 expression both in the tumour and stroma (T+/S+), just in the tumour (T+/S−), just in the stroma (T−/S+) or not present in either of these compartments (T−/S−).</p

Nuclear Snail1 protein expression in colon carcinomas.

<p>Expression of Snail protein was determined as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005595#s4" target="_blank">Methods</a> in samples corresponding to colon carcinomas using MAb EC3. Micrographs of several representative stained sections are shown. Panels A–E corresponded to tumours considered positive only in the stroma; panel F, just in the tumour, and panels G–P; in both compartments. The arrow in panel H labels a cell that cannot be clearly classified as tumoral or stromal. In panel O the arrow points at a cell entering a vessel. Bars indicate magnification.</p

Specific survival of stage I, II and III colon tumour patients according to Snail1 expression in the stroma.

<p>The presence of Snail1 in the stroma of stage I, II and III tumours is represented as continuous lines; dotted lines correspond to stroma-negative tumours. In the lower left panel, expression of Snail1 in the stroma was considered as low or high according to the criteria indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0005595#s4" target="_blank">Methods</a>. The significance is indicated in each category.</p

Characteristics of 162 patients with colorectal cancer.

<p>Characteristics of 162 patients with colorectal cancer.</p

Kaplan-Meier specific survival curves for colon carcinoma patients according to Snail1 expression in the stroma.

<p>Discontinuous line represents negative Snail1 immunoreactivity; continuous line, Snail1 positive immunoreactivity. The graphics compare tumours where expression of Snail1 was observed in the stroma with respect to negative ones, regardless of the immunoreactivity in the tumour (left); and with only reactivity in the stroma with respect to negative biopsies (right). The p values are indicated.</p

Representative high-magnification photomicrographs showing double immunofluorescence for NAAA, CD19, CD3 and CD14 in order to characterize the immune cells in the mucosa infiltrate of UC patients.

<p>NAAA immunofluorescence was observed in CD19+ B lymphocytes (A–C), CD3+ T lymphocytes (D–F) and CD14+ macrophages (G–I).</p

Schematic drawings that hypothesize the normal pattern of NAE-PPARα activity (A), and the pro- and anti-inflammatory NAE-PPARα signaling that may occur in the colonic epithelial cells of active UC at disease onset (B) and after a putative treatment with 5-ASA and/or glucocorticoids (C).

<p>Schematic drawings that hypothesize the normal pattern of NAE-PPARα activity (A), and the pro- and anti-inflammatory NAE-PPARα signaling that may occur in the colonic epithelial cells of active UC at disease onset (B) and after a putative treatment with 5-ASA and/or glucocorticoids (C).</p

Primers sequences used for RT-PCR.

<p>Primers sequences used for RT-PCR.</p

Analysis of the number of FAAH-ir cells per area (µm²) in the lamina propria of acute and quiescent (5-ASA and corticoid-treated) UC patients compared to control ones.

<p>A–F: Representative high-magnification photomicrographs showing FAAH immunostaining in the lamina propria. G: Acute UC at disease onset was associated with a dramatic increase in the number of FAAH-ir cells in the infiltrate of the lamina propria. The number of FAAH-ir cells was significantly dropped after treatment, but do not reach control levels. Mann-Whitney U and Wilcoxon tests (N = 15–22): ***<i>P</i><0.001 <i>versus</i> control group; ^###<i>P</i><0.001 <i>versus</i> acute UC group.</p

Representative photomicrographs and densitometrical quantification of PPARα (A–D), NAAA (E–H), NAPE-PLD (I–L) and FAAH (M–P) immunoreactivity in human healthy (control; A, E, I, M), active UC (B, F, J, N) and quiescent UC (C, G, K, O) colonic epithelium depending on treatment.

<p>Active UC at disease onset showed a decrease in PPARα and NAPE-PLD immunoreactivity, but an increase in NAAA immunoreactivity. PPARα immunoreactivity showed significant differences depending on treatment. Treatment with 5-ASA in UC patients restored completely PPARα and NAPE-PLD protein levels to control ones. However, PPARα expression dropped again in UC patients treated with 5-ASA and glucocorticoids, and 5-ASA, glucocorticoids and immunomodulators compared to control group. In contrasts, treatment with 5-ASA and glucocorticoids increased NAPE-PLD immunoreactivity. No changes were observed in FAAH immunoreactivity in the epithelium of control, active (untreated) UC and quiescent (treated) UC patients. Mann-Whitney U and Wilcoxon tests (N = 22–24): *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 versus control group; ^#<i>P</i><0.05, ^##<i>P</i><0.01 <i>versus</i> UC group; ^&<i>P</i><0.05 <i>versus</i> 5-ASA-treated quiescent UC group.</p