Article Figures & Data
Figures
- Figure 1.
Light micrographs showing periodic acid-Schiff staining of glomeruli counterstained with hematoxylin from Akita (Ins2+/C96Y/B2R+/+) and Akita (Ins2+/C96Y/B2R-/-) genotypes. B2R, bradykinin 2 receptor.
- Figure 2.
Modular organization of ShcA isoforms. CH, collagen-homologous; PTB, phosphotyrosine binding domain; SH, Src homology.
- Figure 3.
Periodic acid-Schiff (PAS) staining of kidney sections. The Ins2 mutation alone increases accumulation of PAS-positive matrix at 12 months of age, which is enhanced by the absence of the bradykinin 2 receptor. The p66 null mutation attenuates renal histological changes at corresponding intervals: nodular PAS-positive matrix (black arrows in Akita and double mutant Akita [DMA]), mesangiolysis (yellow arrows in Akita and DMA), tubular dilation (blue arrow in DMA), and microaneurysm (arrowhead in DMA). The scale bar is 10 μm. The insert shows the histologic analysis of glomerulosclerosis by semiquantitative morphometric analysis. Results are presented as mean ± standard deviation; n=5 in each group; P<0.01 Akita (Ins2+/C96Y) vs p66 knockout (KO) Akita (p66-/-/Ins2+/C96Y); P<0.001 DMA (Ins2+/C96Y/B2R-/-) vs triple mutant Akita (TMA) (p66-/-/Ins2+/C96Y/B2R-/-). WT, wild type.
- Figure 4.
The p66 null mutation attenuates urine albumin excretion. Results are presented as mean ± standard deviation; n=5 in each group. P<0.001 Akita (Ins2+/C96Y) vs p66 knockout (KO) Akita (p66-/-/Ins2+/C96Y); P<0.001 double mutant Akita (DMA) (Ins2+/C96Y/B2R-/-) vs triple mutant Akita (TMA) (p66-/-/Ins2+/C96Y/B2R-/-).
- Figure 5.
Trichrome staining of the renal cortex. Double mutant Akita (DMA) mice (Ins2+/C96Y/B2R-/-) show increased interstitial fibrosis at 12 months of age, whereas the p66 null mutation attenuates interstitial fibrosis at corresponding intervals. The scale bar is 50 μm. The insert shows the histologic analysis of interstitial fibrosis assessed by semiquantitative evaluation. Results are presented as mean ± standard deviation; n=5 in each group. P<0.001 Akita vs p66 knockout (KO) Akita; P<0.001 DMA vs triple mutant Akita (TMA) (p66-/-/Ins2+/C96Y/B2R-/-). WT, wild type.
- Figure 6.
Podocyte number/glomerulus. WT1 (Wilms tumor 1) is expressed in the podocyte nucleus only. Marker-expressing nuclei were counted in the kidneys of Akita mice. Results are presented as mean ± standard deviation; n=5 in each group. Thirty glomeruli from each mouse were analyzed; P<0.001 p66 knockout (KO) Akita (p66-/-/Ins2+/C96Y) vs Akita (Ins2+/C96Y); P<0.001 double mutant Akita (DMA) (Ins2+/C96Y/B2R-/-) vs triple mutant Akita (TMA) (p66-/-/Ins2+/C96Y/B2R-/-). Confocal images of glomeruli show the podocyte-specific marker synaptopodin (red), podocyte nuclear marker WT1 (green), and nuclear counterstain DAPI (blue) in kidney sections at 12 months of age. Merge shows the expression of synaptopodin, WT1, and DAPI.
- Figure 7.
(A) Representative immunoblot analysis of ShcA isoforms in conditionally immortalized differentiated human podocytes (CIDHPs) stably transfected with p66 shRNA. (B) CIDHP/p66 shRNA shows inhibition of HG-induced reactive oxygen species (ROS) metabolism. (C) Quantitative analysis of ROS. (D) CIDHP/p66 shRNA shows inhibition of HG-induced apoptosis signal, *P<0.01. EV-CIDHP, empty vector-transduced conditionally immortalized differentiated human podocytes; HG, high glucose (40 mM); Manni, mannitol; NG, normal glucose (5 mM); OD, optical density.
- Figure 8.
Representative immunoblot analysis of sirtuin 1 (SIRT1) deacetylase and acetylation of lysine-382 of p53 protein. SIRT1 expression is downregulated in Akita diabetic mice but restored in p66 knockout mice. Similarly, the expression of acetyl p53 is upregulated in Akita diabetic mice but returns to normal expression levels in p66 knockout mice. β-actin was used as a loading control. KO, knockout; WT, wild type.
