Kidney phenotype at 30 weeks of age.

<p>Values are means ± SEM; n, number of mice;</p>*<p>P<0.05 between diabetic and control animals.</p

Gene expression in cardiac tissue.

<p>Cardiac gene expression at 10 weeks of age; mRNA levels in diabetic mice relative to controls (assigned as 1.0) normalized to β-actin mRNA level. Values are means ± SE; n, number of mice; TGFβ, transforming growth factorβ; MCP-1, monocyte chemoatactic protein-1; IL1β, interleukin 1β; TNFα, tumor necrosis factor-α; CTGF, connective tissue growth factor.</p>*<p>P = 0.001 between diabetic and control animals.</p

Cardiac injury in male Akita mice.

<p>(A–B) Panels A and B show representative photomicrograph of heart sections from controls (A) and diabetic mice (B) at 10 weeks of age. Examination of Akita mice hearts revealed mild interstitial fibrosis (green) and focal inflammatory infiltrates (arrows). Tissue sections were stained with Masson trichrome and the magnification was 400×. (C) Expression of mRNA for β-myosin heavy chain (β-MHC) was enhanced in 10-week old Akita mice. Levels of mRNA were expressed relative to controls (assigned as 1.0) and normalized to β-actin mRNA level. Black bars = control (n = 5); white bars = Akita (n = 5). Data are mean ± SEM. *P<0.05 vs. age-matched controls.</p

Fasting blood glucose levels and urine output in male Akita mice.

<p>(A) FVB/NJ-<i>Ins2</i>^+/C96Y mice developed sustained hyperglycemia by 4 weeks of age, which persisted throughout the duration of the study. (B) Urine output in FVB/NJ-<i>Ins2</i>^+/C96Y was significantly increased by 8 weeks of age and further increased during the study period. Black bars = control (n = 6 to 8); white bars = Akita (n = 6 to 8). Data are mean ± SEM. *P<0.05 †P<0.001 or **P<0.01 vs. age-matched controls.</p

GBM and proximal tubule basement membrane thickness at 30 weeks of age.

<p>Values are means ± SEM; n, number of mice.</p

Podocyte injury in normoalbuminuric diabetic Akita mice.

<p>(A) Albuminuria was not significantly different in Akita mice and controls at 4 weeks of age. (B) In contrast, urinary nephrin excretion was increased in normoalbuminuric 4-week old Akita mice compared to control animals. (C) There was a significant increase in podocyte apoptosis in Akita mice compared to age-matched controls without a change in podocyte number. Black bars = control (n = 6 to 8); white bars = Akita (n = 6 to 8). Data are mean ± SEM. *P<0.05 or †P<0.001 vs. age-matched controls.</p

Glomerular histopathology of diabetic Akita mice.

<p>Photomicrographs of mouse kidneys were taken using tissue samples prepared from mice at 30-weeks of age. (A) A representative picture from a control mouse (score of 0). (B–D) Moderate mesangial expansion in glomeruli from Akita mice (score of 2). (E) A representative picture of glomerular ultrastructure from a control mouse. (F) FP effacement was not detected by electron microscopy in diabetic Akita mice at 30-weeks of age. Light microscopic sections were stained with Periodic acid-Schiff (PAS) and the magnification was 400×.</p

Renal phenotype of male Akita mice by genetic background.

<p>NR: not reported, ND: Not detected,</p>*<p>Compared to strain matched non-diabetic controls,</p>†<p>Animal models of diabetic complications consortium (ADMCC) criteria: >10-fold increase in albuminuria, >50% decrease in GFR over the lifetime of the animal, Advanced mesangial expansion, Arteriolar hylanosis, >50% increase in GBM thickness, Tubulointerstitial fibrosis,</p>**<p>variable (see text for discussion),</p>‡<p>Increased BUN at 30–40 weeks of age, References: <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Breyer1" target="_blank">[3]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Gurley1" target="_blank">[4]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Gurley2" target="_blank">[5]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Susztak1" target="_blank">[13]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Brosius1" target="_blank">[30]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Hong1" target="_blank">[35]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033942#pone.0033942-Haseyama1" target="_blank">[48]</a>.</p

Echocardiographic analysis.

<p>Echocardiographic analysis was performed at 10 weeks of age. Values are means ± SEM; n, number of mice; IVSW, interventricular septal width; LVDd, left ventricular end diastolic diameter; LVDs, left ventricular end systolic diameter; LVFS, left ventricular fractional shortening; LVm, left ventricular mass; PW, posterior wall thickness.</p>*<p>P<0.001 between diabetic and control animals.</p

Body weight.

<p>Values are means ± SEM; n, number of mice.</p