Mono-dansyl-cadaverine (MDC) fluorescence in isolated rat islets exposed for 6, 12 and 24 h to 0.5 mM PA.

<p>Isolated islets, previously loaded with the fluorescent probe, were deposited on microscope slides by centrifugation with a cytocentrifuge, as further detailed in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036188#s2" target="_blank">Methods</a>.</p

Electron microscopy of isolated rat islets after 6 h incubation with glucose and/or palmitate.

<p><b>A</b>) control; <b>B</b> 25 mM glucose; <b>C</b>) 0.5 mM PA; <b>D</b>) 25 mM glucose+0.5 mM pA (magnification: 10.000×; insert: 38.000×).</p

Immunoblotting of LC-3 protein in INS-1E cells after 2, 6 and 12 h treatment with various concentrations of palmitate (PA).

<p>α-tubulin was used as a control for loading. Methodological details are described in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036188#s2" target="_blank">Methods</a>. The figure is representative of gels obtained from three separate experiments. From these gels, band intensities were evaluated by densitometric analysis and expressed as the mean ± SEM of the ratio between LC3-II and LC3-I bands (lower graphs). * p<0.01 vs. untreated control; § p<0.01 vs. PA 0.5 mM.</p

Cellular localization of LC-3 and cathepsin D immunofluorescence in INS-1E cells after 6 and 24 h treatment with 0.5 and 1.0 mM PA.

<p>Cellular localization of LC-3 and cathepsin D immunofluorescence in INS-1E cells after 6 and 24 h treatment with 0.5 and 1.0 mM PA.</p

Figure 10

<p><b>A</b>) Ultrastructural morphometric analysis of beta-cell death in human islets after 48 h exposure to 0.5 mM palmitic acid ±10 ng/ml rapamycin. C = control; P = 0.5 mM palmitic acid; P+R = 0.5 mM palmitic acid plus 10 ng/ml rapamycin. Mean of 3–4 samples ± SEM. Statistical analysis (Bonferroni's test): * p<0.05 vs control and vs palmitate+rapamycin; # p>0.05 vs palmitate. <b>B</b>) Electron microscopy of isolated human islets in control conditions (left) and after 48 h incubation with 0.5 mM PA (center) or 0.5 mM PA+10 ng/ml rapamycin (right). (magnification: 10.000×).</p

Electron microscopy of isolated human islets after 24 h incubation with glucose and/or palmitate.

<p><b>A</b>) control; <b>B</b> 25 mM glucose; <b>C</b>) 0.5 mM PA; <b>D</b>) 25 mM glucose+0.5 mM pA (magnification: 10.000×; insert: 38.000×).</p

Electron microscopy of isolated rat islets after 24 h incubation with glucose and/or palmitate.

<p><b>A</b>) control; <b>B</b> 25 mM glucose; <b>C</b>) 0.5 mM PA; <b>D</b>) 25 mM glucose+0.5 mM pA (magnification: 10.000×; insert: 38.000×).</p

Electron microscopy of isolated human islets after 6 h incubation with glucose and/or palmitate.

<p><b>A</b>) control; <b>B</b> 25 mM glucose; <b>C</b>) 0.5 mM PA; <b>D</b>) 25 mM glucose+0.5 mM pA (magnification: 10.000×; insert: 38.000×).</p

Representative image (left panel) of a pancreatic cell in T2D samples containing both amylase (black arrows) and insulin (white arrows) granules, with adjacent cells of the immune system.

<p>The image has been reconstructed from several pictures at 10,000x final magnification. Enlargments are provided in the right panels. Mc: macrophage, Lymph: lymphocyte, Ms: mast cell.</p

Electron microscopy images (magnification 10,000x) of pancreatic cells containing both zymogen-like (black arrows) and insulin-like (white arrows) granules in the four cases with T2D.

<p>Electron microscopy images (magnification 10,000x) of pancreatic cells containing both zymogen-like (black arrows) and insulin-like (white arrows) granules in the four cases with T2D.</p