Histological picture of a laser microdissected biopsy.

<p>Example of laser-microdissection pre- (A) and post- (B) cut, showing that the glomerular tuft can be separated without any tubulo-interstitial contamination (Ematoxilin staining).</p

Real-Time quantification of ClC-5 in laser-microdissected biopsies of NIDDM and controls.

<p>mRNA level of ClC-5 in laser-microdissected glomeruli from NIDDM cases and controls. ClC-5 mRNA levels were significantly higher in NIDDM glomeruli than in control glomeruli (p<0.002). The relative expression levels of the gene of interest was obtained by dividing the gene’s expression level by the mean level of the different housekeeping genes. The relative levels of the controls were set to one.</p

Immunolocalization of ClC-5 in ultrastucture of MG biopsies.

<p>A. Transmission electron microscopy (TEM) of MG biopsy revealed small vesicles in the secondary foot processes of podocytes (direct magnification 9000×). B–C. Ultrastructural immunolocalization of ClC-5 in MG biopsy. Arrows indicate some of the gold particles located in podocytes (direct magnification 40000×).</p

Patients’ clinical details.

<p>Note: Conversion factors for units: serum creatinine in mg/dl to mol/L, x88.4.</p

Double staining for ClC-5 and WT1 in MG and control biopsies.

<p>Co-localization of ClC-5 (cytoplasmatic brown stain) and WT1 (nuclear blu-grey stain) in control (C) and in MG (A) glomeruli to identify podocytes (400×). B. Oil image immersion of a detail of panel A (1000×).</p

Immunoistochemical analysis in MG biopsies.

<p>A. Glomerular ClC-5 immunostaining in MG patient (400×). B. ClC-5 staining in a control glomerulus (400×). Black arrows indicate cytoplasmatic ClC-5 staining (400×). C. Proximal tubular cells used for internal control purposes showed ClC-5 apical and sub-apical staining (200×). D. ClC-5 quantification by morphometric analysis on MG and control glomeruli. ClC-5 signal was significantly higher in MG than in control samples (p<0.01). The quantity was expressed as the mean of the area covered by pixels, as a percentage. The average of the morphometric data in each group of biopsies is reported. E. Absence of glomerular ClC-5 staining in a patient with Dent’s disease (400×). F. Negative control obtained by incubating the sample with no primary antibody or with nonimmune rabbit IgG (400×).</p

Model characteristics at 15 months of follow up.

<p>Model characteristics at 15 months of follow up.</p

Early stages of atherogenic DM leads to renal damage.

<p>A: Representative illustration of PAS stained glomeruli from a DM+ATH pig, showing mesangial proliferation and matrix expansion with capillary loops lying around the mesangium as a corona, reminiscent of a beginning Kimmelstiel-Wilson nodule (left panel; thin black arrow). Dilated capillary loops with red cell fragments show intense PAI-1 staining on consecutive slides (right panel; thick black arrow). B: Mesangial expansion index in Controls (n = 7), ATH (n = 5) and DM+ATH (n = 5) pigs. C. Electron microscopy images illustrating a normal GBM architecture (left panel; thick arrow) of the Controls pig. In ATH, there is slight effacement of the podocyte pedicles (middle panel; thick arrow). In DM+ATH, marked lipid deposits were found (right panel). Data are shown as mean ± SEM. *P<0.05 compared to Controls or ATH pigs. Original magnification of A: x400 and C: x8000.</p

No difference in renal vWf and VEGF-A expression.

<p>A. Representative illustrations of kidney sections stained with endothelial marker vWF (arrow: glomerulus; arrowhead: peritubular area) in Controls, ATH, and DM+ATH pigs. B. Representative images of kidney sections stained with VEGF in Controls, ATH, and DM+ATH pigs showing expression in podocytes (arrow head), parietal epithelial cells (thin arrow) and tubuli (asterix). Original magnification of A: x 200 and B: x400.</p

Correlation between capillary tortuosity and Angpt2/Angpt1balance and creatinine levels.

<p>Scatter plot showing the correlation of renal protein expression of Angpt1(A), Angpt2 (B), Angpt2/Angpt1ratio (C).</p