Changes in mRNA cytokine expression levels in mouse abdominal fat tissue at different time points post IP injection of AuNPs.

<p>mRNA levels of (A) CD68, (B) TNFα and (C) IL-6 in abdominal fat at different time points post AuNPs injection. Results are expressed as mean ± SE, n = 4–8 in each group. *P<0.05, compared with Control group. CON, control; Au, AuNPs. The differences among groups were analysed using one-way ANOVA, followed by <i>post hoc</i> Fisher's Least Significance Difference tests.</p

AuNP distribution within mouse liver following IP injection.

<p>AuNPs in (A) the liver connective tissue and (B) a 5 µm capillary within the liver adipose tissue (Mag 5.00K x), and in (C) a Kupffer cell within the sinusoid of the liver (Mag 4.19K x).</p

Additional file 1: of FXR expression is associated with dysregulated glucose and lipid levels in the offspring kidney induced by maternal obesity

Supplementary data. (DOCX 96 kb

Abdominal adipose tissue distribution of AuNPs following intraperitoneal injection into mice.

<p>Scanning electron microscopy (SEM) images of abdominal adipose tissue in (A) control mouse and (B) AuNP-injected mouse at 24 h (Mag 2.01K x). (C, D) Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) images of abdominal adipose tissue at 24 h. (C) displaying the carbon present throughout the tissue sample, of medium to low signal intensity, (D) displays the gold present throughout the tissue sample, concentrated around the periphery of a portion of tissue within the section, of medium to high intensity. The strength of the signal transmitted for a selected element is represented by a corresponding colour, blue signifying the lowest signal strength and red the highest.</p

Metabolic and physiological parameters of UUO mice treated with Telmisartan or PXS64.

<p>Metabolic and physiological parameters of UUO mice treated with Telmisartan or PXS64.</p

High glucose and TGF-β1 induced Nox4 expression.

<p>A) Real time PCR showing increased TGF β1 mRNA expression in high glucose treated HK2 cells. B) ELISA showing TGF β1 levels in pg/ml of protein produced following exposure of HK2 cells to high glucose. Real time PCR showing Nox4 mRNA expression following exposure to high glucose (C) or TGF-β1 (D). HK2 were incubated for 24 hrs with 5 mM glucose media (control), 30 mM D glucose or TGF-β1 (2 ng/ml). Data is normalised by glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and expressed as mean ± SEM; n = 3. <i>*P</i><0.05 vs. Control.</p

PXS64 inhibited TGF-β1-induced fibrotic and inflammatory markers in HK2 cells.

<p>HK2 cells exposed to 100ng/ml latent TGF-β1 showed a significant increase in fibronectin and collagen IV mRNA (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116888#pone.0116888.g002" target="_blank">Fig. 2A and B</a>) and protein (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116888#pone.0116888.g002" target="_blank">Fig. 2D and E</a>) expression, which were suppressed when co-exposed to PXS-64. These cells had increased mRNA expression of MCP-1 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116888#pone.0116888.g002" target="_blank">Fig. 2C</a>), which was translated to increased secretory MCP-1 in the supernatant (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116888#pone.0116888.g002" target="_blank">Fig. 2F</a>). There were no difference mRNA and protein expression of the fibrotic and inflammatory markers s in HK2 cells under basal conditions or when cells were exposed to PXS64 alone (*<i>P</i> < 0.05, ** p<0.01 vs. appropriate control) Results are expressed as mean ± SEM, n = 4</p

Nox4 expression and ROS production <i>in vivo</i>.

<p>Immunohistochemistry demonstrating that Nox4, nitrotyrosine and 8-OHdG protein expression are highly expressed in STZ-induced diabetic C57BL/6J mice (B, E and H respectively) compared to controls (A, D and G). Plumbagin (2 mg/kg/day) administration blocked diabetic induced Nox4, nitrotyrosine and 8-OHdG expressions (C, F and I). Proportion of area immunostained for Nox4, nitrotyrosine and 8-OHdG are shown (J, K, L). STZ and plumbagin were administered as described in the Material and Methods. Negative IgG was performed to confirm staining specificity (not shown). Magnification x 400.</p

Reduction in cellular infiltration and inflammatory markers in UUO kidneys exposed to telmisartan or PXS64.

<p>Untreated UUO kidneys showed increased F4/80, CD68 and CD45 positively stained cells as compared to the sham operated control animals. PXS64 significantly reduced F4/80 and CD45 positive stained cells (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116888#pone.0116888.g006" target="_blank">Fig. 6C and E</a>) with a trend to a reduction in CD68 cells, although this was not statistically significant (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0116888#pone.0116888.g006" target="_blank">Fig. 6D</a>). Telmisartan treated kidneys showed a reduction in F4/80 positive cells but no difference in CD45 or CD68 stained cells, suggesting a differential action of PXS64 and telmisartan in modifying cellular infiltration. Results are presented as mean showed (n = 8, *P < 0.05 vs. UUO, ** P < 0.01 vs. UUO). Magnification x 400.</p

Localisation of AuNPs within specific mouse tissues following IP injection.

<p>Scanning electron microscopy (SEM) images of AuNPs in adipose tissue (A, B, C) and liver sections (D, E, F) at 1 h (A, D), 24 h (B, E), and 72 h (C, F) post AuNP injection (Mag 2.01K x).</p