Baseline parameters are comparable between wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice.

<p>The number of circulating leukocytes (<b>A</b>) and neutrophils (<b>B</b>) in the peripheral blood of wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice were quantitated. Bone marrow leukocytes (<b>C</b>) and neutrophils (<b>D</b>) isolated from wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice were quantitated. N ≥ 8 per group. Baseline cell surface expression of L-selectin (<b>E</b>), and PMA-induced L-selectin shedding (<b>F</b>) and respiratory burst (<b>G</b>) was measured and compared between wildtype and <i>Gpr43</i>^{<i>−/−</i>}bone marrow neutrophils. N ≥ 4 mice per group or independent <i>in vitro</i> experiments performed in duplicates, N.S. denote not statistical significant, <i>t</i> test.</p

Acetate reduces neutrophil migration via a mechanism mediated by GPR43.

<p>The total number of leukocytes in the peritoneal cavity was quantified in wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice at 24 h following sham or cecal ligation and puncture (CLP; (<b>A</b>)), or at 2 h after saline, fMLP or fMLP + acetate (<b>B</b>). The peritoneal neutrophil numbers of acetate-treated wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice at 2 h after saline or fMLP treatment were also measured (<b>C</b>). N ≥ 3 mice per group, **<i>p</i> < 0.01, *<i>p</i> < 0.05 vs corresponding Sham or Saline group, <i>t</i> test. ##<i>p</i> < 0.01, #<i>p</i> < 0.05 vs treated Wildtype, <i>t</i> test. <i>&p <</i> 0.05 vs fMLP-treated Wildtype, <i>t</i> test. N.S denotes not statistically significant vs fMLP-treated <i>Gpr43</i>^{<i>−/−</i>}.</p

Exacerbated intestinal inflammation in <i>Gpr43</i>^{<i>−/−</i>}mice in response to LPS.

<p>(<b>A</b>) Representative histological images of jejunum of wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice following 4 h of saline or LPS challenge. Neutrophils are denoted by the black arrowheads. Scale bar = 100 μm. (<b>B</b>) Histological quantification of neutrophils in the (<b>i</b>) duodenum, (<b>ii</b>) jejunum and (<b>iii</b>) ileum of wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice following 4 h of saline or LPS challenge. N ≥ 5 mice per group, **<i>p</i> < 0.01, *<i>p</i> < 0.05 vs corresponding Saline group, <i>t</i> test. #<i>p</i> < 0.05 vs Wildtype, <i>t</i> test.</p

GPR43-deficient neutrophils display slow rolling following 4 h of LPS challenge.

<p>Intravital microscopy was utilised to visualise and quantitate the number of intravascular neutrophils that were rolling (<b>A</b>), and their rolling velocity (<b>B</b>) or adherent (<b>C</b>) following 4 h of saline or LPS challenge in wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice. N ≥ 5 mice per group, ***<i>p</i> < 0.001 vs corresponding Saline group, <i>t</i> test. # <i>p</i> < 0.05 vs Wildtype, <i>t</i> test.</p

GPR43 deficiency induces migration, accelerated neutrophil rolling and adhesion following 1 h of LPS challenge.

<p>Neutrophils were isolated from the bone marrow of wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice, and analysed for chemotaxis towards increasing concentration of CXCL1 (<b>A</b>) and LPS (<b>B</b>). N ≥ 4 independent <i>in vitro</i> experiments performed in duplicates, ***<i>p</i> < 0.001, *<i>p</i> < 0.05 vs Wildtype, 2-way ANOVA. Intravital microscopy was utilised to visualise and quantitate the number of intravascular neutrophils that were rolling (<b>C</b>) or adherent (<b>D</b>) following 1 h of saline or LPS challenge in wildtype and <i>Gpr43</i>^{<i>−/−</i>}mice. N ≥ 5 mice per group, ***<i>p</i> < 0.001, **<i>p</i> < 0.01 vs corresponding Saline group, <i>t</i> test. #<i>p</i> < 0.05 vs Wildtype, <i>t</i> test.</p

Enhanced migratory behaviour of neutrophils is maintained after microbiota transfer.

<p>Neutrophils of GF mice reconstituted with microbiota from SPF mice fed on Ctrl or NF diet were analyzed for chemotaxis towards increasing concentration of CXCL1 (<b>A</b>) and fMLP (<b>B</b>). N ≥ 4 independent <i>in vitro</i> experiments performed in duplicates, **<i>p</i> < 0.01, *<i>p</i> < 0.05 vs Ctrl diet, 2-way ANOVA.</p