Animal characteristics for NOD diabetic mice (Diabetic), NOD diabetic mice treated with insulin (Diabetic+INS) and NOR control mice (Control).

<p>Animal characteristics for NOD diabetic mice (Diabetic), NOD diabetic mice treated with insulin (Diabetic+INS) and NOR control mice (Control).</p

Blood pressure and kidney function parameters in non-treated non-obese diabetic mice (Diabetic), NOD-insulin treated mice (Diabetic+INS) and non-obese resistant non-diabetic mice (Control).

<p>A: At the end of the early stage study, systolic and diastolic blood pressure measured by tail-cuff method in Diabetic, Diabetic+INS and Control mice. B: Urinary albumin/creatinine ratio of Diabetic, Diabetic+INS and Control mice at both study stages. C: Glomerular filtration rate measured by inulin in Diabetic, Diabetic+INS and Control mice studied at early stage of diabetes. *<i>P≤0.05.</i></p

Morphometric analysis for NOD diabetic mice (Diabetic), NOD diabetic mice treated with insulin (NOD+INSULIN) and NOR control mice (Control).

<p>Morphometric analysis for NOD diabetic mice (Diabetic), NOD diabetic mice treated with insulin (NOD+INSULIN) and NOR control mice (Control).</p

ACE2 activity in serum (A), urine (B) and kidney cortex (C) from non-treated non-obese diabetic mice (Diabetic), NOD-insulin treated mice (Diabetic+INS) and non-obese resistant non-diabetic mice (Control) (A, B, C).

<p>ACE2 protein expression measured by western blot of protein preparations from kidney cortex lysates in Diabetic, Diabetic+INSand Control mice (D). Immunoblot analysis of ACE2 in kidney cortex tissue (E). Shown is representative immunoblot of ACE2 protein in kidney cortex from Control, Diabetic, and Diabetic+INS mice, at early and late stage of diabetes. *<i>P≤0.05.</i></p

Effect of Insulin on ACE2 Activity and Kidney Function in the Non-Obese Diabetic Mouse - Figure 2

<p><b>A. Representative micrographs of periodic acid–Schiff (PAS)-stained kidney sections.</b> Upper panel: Diabetic, Diabetic-INS early stage of diabetes and Control mice. B: Lower panel: Diabetic, Diabetic-INS late stage of diabetes and Control mice. <b>B. Podocyte number in Diabetic, Diabetic-INS, and Control mice.</b> Representative photomicrograph depicting glomerular WT-1 staining in glomeruli from Diabetic, Diabetic-INS mice at late stage of diabetes and Control mice. Original magnification X400. <b>C. Ultrastructural studies in non-treated non-obese diabetic mice, Diabetic-insulin treated mice and Controls.</b> Upper panel: Diabetic, Diabetic-INS early stage of diabetes and Control mice. Lower panel: Diabetic, Diabetic-INS late stage of diabetes and Control mice. Glomeruli from late stage NOD Diabetic mice (lower middle panel) showed increased mesangial expansion due to increased mesangial matrix increase (see asterisks) as compared with Control mice. Insulin administration prevented mesangial expansion in NOD Diabetic mice. Original magnification, X4600.</p

Loss of ARID1A expression is associated with more aggressive UBC and with patient outcome.

<p>ARID1A expression was assessed by IHC on tissue microarrays. Patients (n = 84) were followed-up as indicated in Methods and classified as having “recurred”, “progressed”, or being free of disease. Patients with high ARID1A-expresssing tumors display a lower risk of recurrence and a higher risk of progression indicating a more aggressive clinical course.</p

<i>ARID1A</i> mutations and expression in UBC.

<p><i>Panel A</i>. A G>C transversion identified through Solexa resequencing, confirmed by Sanger sequencing of independent PCR products, leading to a predicted Q2210H substitution in VMCUB-3 cells. <i>Panel B</i>. Western blotting analysis in a panel of UBC cell lines identifies a subset with undetectable expression, including VMCUB-3. mRNA expression was analyzed by RT-qPCR; results are shown as values normalized with respect to the housekeeping gene <i>HPRT</i>. <i>Panel C</i>. A C>T mutation in codon 403, leading to a premature stop codon, was identified in a primary T1G3 tumor. The mutation was absent from matched normal leukocyte DNA. Lack of protein expression in the corresponding tumor tissue was confirmed using immunohistochemistry. The red arrowhead points to a tumor cell lacking ARID1A staining, whereas the black arrowhead indicates a positive stromal cell. For comparison, a TaG1 tumor with wild type <i>ARID1A</i> sequence is shown.</p

Kaplan-Meier survival curves corresponding to failures in superficial (A, B) and invasive (C, D) tumors for specified prognostic endpoints.

<p>Dashed curves: patients with tumors positive for COX2 protein staining; solid curves: patients with tumors negative for COX2 protein staining. Significance values from two-sided logrank test.</p

Effects of <i>ARID1A</i> knockdown in UBC cell lines.

<p><i>Panel A</i>. <i>ARID1A</i> was knocked-down using three different shRNAs in the RT112 and VMCUB-3 cells. The knock-down was efficient at both the protein and mRNA levels. The bars represent the relative quantification of ARID1A mRNA levels taking non-targeting shRNA interfered cells as controls. <i>Panel B</i>. The quantification colony formation is shown, with error intervals of results from triplicate experiments denoting SEM. In RT112 cells, ARID1A knockdown was associated with reduced colony formation. By contrast, no major effects were observed in VMCUB-3 cells harboring an <i>ARID1A</i> mutation. Representative morphological changes in cells interfered with control shNT (scrambled shRNA) and with one of the shRNAs targeting <i>ARID1A</i> are shown.</p

Characteristics of the patients from whom fresh tumor was used for <i>ARID1A</i> sequence analysis.

<p>Characteristics of the patients from whom fresh tumor was used for <i>ARID1A</i> sequence analysis.</p