Evidences for Expression and Location of ANGPTL8 in Human Adipose Tissue

Figures

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  Figure 1. Plasma circulating levels of C-terminal 139–198 form of ANGPTL8. Data was expressed as mean ± the standard error of the mean (SEM), and the values in the non-obese women were 0.63 ± 0.10 ng/mL, n = 11 and obese women 0.64 ± 0.04 ng/mL, n = 33. In non-obese men 0.82 ± 0.09, n = 10 versus 0.83 ± 0.10 ng/mL in obese men, n = 14. The Student’s t-test analysis in non-obese patients (women vs. men) was p = 0.0763, and in obese patients (women vs. men) p = 0.176 (n.s.).

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  Figure 2. Detection of ANGPTL8 in human visceral adipose tissue (VAT). Representative images from the immunofluorescence reaction for positive ANGPTL8 (in red) in adipose cells of patients, (A, non-obese women, n = 4), (B, obese women, n = 4), (C, non-obese men, n = 4), (D, obese men, n = 4). Scale bar: 60 μm. (E) Western blotting analysing the expression of ANGPTL8 in human VAT (n = 6), ERK1/2 was used to confirm equal loading of samples. (F) Plot showing the relative ANGPTL8 expression as arbitrary units (AUs).

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  Figure 3. Co-existence of perilipin-1 and ANGPTL8 in human VAT. Representative images showing the double immunofluorescence reaction for positive anti-perilipin-1 (in green), ANGPTL8 (in red) and the merge in adipose cells of patients (in yellow), (A, non-obese women, n = 4), (B, obese women, n = 4), (C, non-obese men, n = 4), (D, obese men, n = 4). Scale bar: 63 μm.

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  Figure 4. Expression of ANGPTL8 in human VAT. Real time polymerase chain reaction (RT-PCR) analysing ANGPTL8 relative gene expression in human VAT (non-obese women, n = 6; non-obese men, n = 5; obese women, n = 6; obese men, n = 5) calculated by normalisation to Ribosomal Protein L19 (RPL19) and expressed as arbitrary units (AUs).

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  Figure 5. Representative images showing the immunofluorescence reaction for the endothelial cells’ marker anti-CD34 in adipose tissue (micrographs A, D, G, J, in green). The micrographs B, E, H, K present the in situ hybridisation for ANGPTL8 in VAT (ANGPTL8 mRNA, in red). The micrographs C, F, I, and, L are the merge of both images, (non-obese women, n = 4), (obese women, n = 4), (non-obese men, n = 4), (obese men, n = 4). Scale bar: 60 μm.