Composition of the diets.

<p>Composition of the diets.</p

Fibrosis is increased in the liver of <i>E</i>. <i>cloacae</i> -treated mice.

<p>(A), (B), and (C) H&E staining; (D), (E) and (F) overexposed anti -smooth muscle actin stainings of the frozen liver tissues. H&E staining shows more ballooning and an increase in lipid droplets in HFD controls and <i>E</i>. <i>cloacae</i> -treated mice compared to chow controls. In SMA staining, an increase in <i>E</i>. <i>cloacae</i> -treated mice compared to chow controls and HFD controls can be seen both in blood vessels (discontinuous arrows) and sites outside the vessels (continuous arrows).</p

<i>E</i>.<i>cloacae</i> adminstration decreases the number of leukocytes and increases the size of adipocytes.

<p>(A) and (B) The histological images present the adipose tissues with CD45-staining indicated with arrows. (C) The relative numbers of CD45-positive cells as determined with manual cell counting from the stained tissues, represented as average cell numbers per field of view. (D) The percentages of adipocytes with diameters of 10–20 μm, 20–30 μm and 30–40 μm in each group. 500 cells were randomly selected from each sample. All data are presented as mean ± SD. n was 5-6/group. The statistical significance was set to p<0.05 and the significant differences are presented with lines and * between the groups.</p

<i>E</i>. <i>cloacae</i> administration increases insulin resistance and lipolysis, but also increases adiponectin expression in VAT.

<p>At necropsy (A) visceral fat mass; (B) <i>Adipoq</i> expression; (C) <i>Rela</i> expression; (D) <i>Insr</i> expression; (E) AKT phosphorylation; (F) HSL phosphorylation; (G) <i>Mgll</i> expression; (H) <i>Ccl2</i> expression; (I) <i>Tlr4</i> expression; (J) The relative numbers of CD45-positive cells as determined with manual cell counting from the stained tissues, represented as average cell numbers per field of view. All data are presented as mean ± SD. n was 4-6/group when statistical outliers were excluded. The statistical significance was set to p<0.05 and the significant differences are presented with lines and * between the groups.</p

<i>E</i>. <i>cloacae</i> administration increases TLR5 and IL-22 gene expression.

<p>At necropsy: (A) <i>Tjp1</i> expsession; (B) <i>Tlr5</i> expression; (C) <i>Il1b</i> expression; (D) <i>Rela</i> expression; (E) <i>Ccl2</i> expression; (F) <i>Il22</i> expression. All data are presented as mean ± SD. n was 4-6/group when statistical outliers were excluded. The statistical significance was set to p<0.05 and the significant differences are presented with lines and * between the groups.</p

Summary of the main findings.

<p><i>E</i>. <i>cloacae</i> administration increased adipose tissue hypertrophy and hepatic damage in the HFD fed mice. The subcutaneous adipose tissue of the <i>E</i>. <i>cloacae</i> -treated mice appeared to be partly insulin resistant, and the increased lipolysis and adipocyte hypertrophy led to increased glycerol release. Liver fat accumulation did not increase in response to the <i>E</i>. <i>cloacae</i> treatment, but AST activity measurements and histology revealed hepatic damage in the <i>E</i>. <i>cloacae</i> -treated mice.</p

<i>E</i>. <i>cloacae</i> administration increases subcutaneous adipose tissue mass and reduces insulin receptor β expression in SAT.

<p>(A) Weight gain during the treatment period. (B) Food consumption during the treatment period. (C) Average daily energy intake during the treatment period. At necropsy: (D) SAT mass; (E) <i>Insr</i> expression; (F) <i>Xbp1</i> expression; (G) <i>Ccl</i> expression; (H) <i>Tlr4</i> expression. All data are presented as mean ± SD. n was 4-6/group when statistical outliers were excluded. The statistical significance was set to p<0.05 and the significant differences are presented with lines and * between the groups.</p

Settlement of Joint Marital Property

<p>Correlation of liver fat content and Matsuda index (= insulin sensitivity) with (A) adipose tissue <i>TLR5</i> mRNA and (B) adipose tissue <i>TLR4</i> mRNA in humans (n = 23).</p

Adipocytes as a Link Between Gut Microbiota-Derived Flagellin and Hepatocyte Fat Accumulation - Fig 6

<p><b>(A) The direct and (B) adipocyte-mediated effects of FLG and LPS on the expression of mitochondrial respiratory chain complex subunits in HepG2 cells.</b> The figure presents the expression levels of mitochondrial respiratory chain complex subunits ATP5A (Compl V), UQCRC2 (Compl III), MTCO1 (Compl IV), SDHB (Compl II) and NDUFS8 (Compl I) in response to 4 and 24 hours exposure to FLG and LPS. All expression levels were normalized to GAPDH and the determined intensities of the Western blot bands are presented as arbitrary units. The values are means + SD (n = 3). * indicates a p-value of <0.05.</p

Adipocytes as a Link Between Gut Microbiota-Derived Flagellin and Hepatocyte Fat Accumulation - Fig 5

<p><b>(A) The direct, and (B) adipocyte-mediated effects of FLG and LPS on insulin signaling in HepG2 cells.</b> The figure presents the mRNA fold changes of <i>IRS1</i> in response to 1, 4 and 24 hour treatments (n = 5) and the phosphorylation levels of Akt in response to 30 min, 1 and 4 hour treatments (n = 4). The mRNA results were normalized to <i>β-actin</i> and the fold changes were calculated with the <i>ΔΔC</i>_<i>t</i> method. Akt phosphorylation levels were normalized to GAPDH and the determined intensities of the Western blot bands are presented as arbitrary units. The values are means + SD. * indicates a p-value of <0.05.</p