Neogenin is expressed outside the CNS.

<p><b>A</b>) Neogenin (<i>Neo 1</i>) mRNA expression in the brain (Br), lung (Lu), liver (Li), spleen (Sp) and intestine (In) of WT animals <b>B</b>) Pooled Western blot analysis of four independent animals demonstrating <i>Neo1</i> expression in (Br), lung (Lu), liver (Li), spleen (Sp) and intestine (In) <b>C</b>) Flow-cytometry of murine blood stained with anti-Neo1, anti-CD45 or isotype-matched control antibody (Data are Mean ± SEM, n = 4 per condition).</p

Loss of haematopoietic neogenin decreases acute peritoneal inflammation.

<p>Chimeric animals and controls were injected i.p. with zymosan A (ZyA) and peritoneal lavage obtained after 8 hours <b>A</b>) Cell count within the peritoneal fluid in chimeric animals and controls 8 h after exposure to ZyA <b>B</b>) Myeloperoxidase (MPO) activity in peritoneal lavage <b>C</b>) Protein content in peritoneal lavage of chimeric animals and controls <b>D</b>) Representative histological analysis of the peritoneum, the mesenterial fat and cytospin samples of the peritoneal lavage of chimeric animals and controls 8 hours following intraperitoneal ZyA injection. Sections prepared with hematoxylin-eosin staining (Magnification ×400, insert ×1000). (Data are Mean ± SEM, n = 6 per group, *P<0.05, **P<0.01, ***P<0.001 as indicated).</p

Neogenin enhances acute inflammatory peritonitis.

<p><i>Neo1^−/−</i> and WT animals were injected i.p. with NaCl or 1 mg of zymosan A (ZyA) and peritoneal lavage obtained after 8 hours <b>A</b>) Cell count in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>B</b>) Myeloperoxidase (MPO) activity in peritoneal lavage <b>C</b>) Protein content in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>D</b>) Representative histological analysis of the peritoneum, the mesenterial fat and cytospin samples of the peritoneal lavage in <i>Neo 1^−/−</i> and WT animals 8 hours following intraperitoneal NaCl 0.9% or ZyA injection. Sections prepared with hematoxylin-eosin staining (Magnification ×400, insert ×1000; Data are Mean ± SEM, n = 6 per group, *P<0.05, **P<0.01, ***P<0.001 as indicated).</p

Functional inhibition of neogenin dampens release of inflammatory cytokines <i>in-vivo</i>.

<p>WT animals were injected i.p. with NaCl or 1 mg of zymosan A (ZyA) and subsequently i.v. with either IgG control or anti-Neogenin (Anti-Neo1) antibody (1 µg) and peritoneal lavage obtained after 8 hours <b>A</b>) TNF-α concentration in peritoneal lavage <b>B</b>) IL-1β concentration in peritoneal lavage <b>C</b>) IL-6 concentration in peritoneal lavage <b>D</b>) KC concentration in peritoneal lavage of WT animals injected with either vehicle or Anti-Neo1 (Data are Mean ± SEM, n = 6 per group, *P<0.05, **P<0.01, ***P<0.001 as indicated).</p

Neogenin increases inflammatory cytokine release <i>in-vivo</i>.

<p><i>Neo1^−/−</i> and <i>WT</i> animals were injected i.p. with NaCl or 1 mg of zymosan A (ZyA) and peritoneal lavage obtained after 8 hours <b>A</b>) TNF-α concentration in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>B</b>) IL-1β concentration in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>C</b>) IL-6 concentration in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>D</b>) KC concentration in peritoneal lavage of <i>Neo1^−/−</i> and WT animals 8 hours following intraperitoneal NaCl or ZyA injection (Data are Mean ± SEM, n = 6 per group, *P<0.05, **P<0.01, ***P<0.001 as indicated).</p

Loss of haematopoietic neogenin dampens cytokine release during acute inflammatory peritonitis.

<p>Chimeric animals and controls were injected i.p. with 1 mg of zymosan A (ZyA) and samples taken after 8 hours <b>A</b>) TNF-α concentration in peritoneal lavage <b>B</b>) IL-1β concentration in peritoneal lavage <b>C</b>) IL-6 concentration in peritoneal lavage <b>D</b>) KC concentration in peritoneal lavage of chimeric animals and controls injected with zymosan A (Data are Mean ± SEM, n = 6 per group, *P<0.05, **P<0.01, ***P<0.001 as indicated).</p

Functional inhibition of neogenin dampens acute inflammatory peritonitis.

<p>WT animals were injected i.p. with NaCl or 1 mg of zymosan A (ZyA) and subsequently i.v. with either IgG control or anti-Neogenin (Anti-Neo1) antibody (1 µg) and peritoneal lavage obtained after 8 hours <b>A</b>) Cell count in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>B</b>) Myeloperoxidase (MPO) activity in peritoneal lavage <b>C</b>) Protein content in peritoneal lavage of <i>Neo1^−/−</i> and WT animals <b>D</b>) Representative histological analysis of the peritoneum, the mesenterial fat and cytospin samples of the peritoneal lavage in <i>Neo1^−/−</i> and WT animals 8 hours following intraperitoneal NaCl or ZyA injection. Sections prepared with hematoxylin-eosin staining (Magnification ×400, insert ×1000; Data are Mean ± SEM, n = 6 per group, *P<0.05, **P<0.01, ***P<0.001 as indicated).</p

UNC5B expression outside the CNS.

<p>A) Relative UNC5B mRNA expression in brain, heart, intestine, blood, lung, kidney, liver and spleen in C57Bl/6 mice. B)–E) Flowcytometry of murine leukocytes gated and stained for the leukocyte marker CD45 (PerCP labeled), the macrophage marker F4/80 (APC labeled) and UNC5B (PE labeled). F) Immune fluorescence staining of human PMN was performed using anti-UNC5B and CD45 antibodies as primary and Alexa488-conjugated and CruzFluor™ 594-conjugated as secondary antibodies were used as well as isotype matched control antibodies. (Data are mean ± SEM, n = 4 per condition).</p

UNC5B^+/− animals show reduced PMN infiltration and activity following hepatic IRI.

<p>A) PNM counts per high power field in WT and UNC5B^+/− mice in the stained liver tissue. B) Relative MPO activity in hepatic tissue of WT and UNC5B^+/− animals C) Representative pictures of PMN staining of liver lobes (brown, marked with arrowheads) of WT and UNC5B^+/− mice after IRI and sham operation are shown (magnification 400×). Data are mean ± SEM, n = 6 per group.</p

UNC5B^+/− animals demonstrate reduced hepatic IRI.

<p>WT and UNC5B^+/− animals underwent 30 minutes ischemia and 3 hours of reperfusion (IR) or sham operation before samples were taken. Serum was analysed for A) LDH, B) ALT, C) AST (Data are mean ± SEM, n = 6 per condition). D) Representative pictures of TTC stained liver lobes are shown (n = 4 per condition).</p