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Table 1.

Animal characteristic and kidney function parameters.

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Table 1 Expand

Fig 1.

Kidney function.

Blood pressure (A) and proteinuria (B) at 4, 8 and 12 weeks and GFR (C-E) as assessed by FITC inulin clearance at 8 weeks (C) and by both FITC inulin (D) and creatinine clearance (CrCl, E) at 12 weeks following subtotal nephrectomy or sham surgery, treated with either vehicle or dapagliflozin. When compared with sham nephrectomised rats, animals that had undergone subtotal nephrectomy developed hypertension, heavy proteinuria and reduced GFR. Subtotally nephrectomised animals that had received dapaglaflozin had similar blood pressure and proteinuria when compared with those that had received vehicle with lower GFR at 12 but not 8 weeks post surgery when assessed by FITC inulin but not by creatinine clearance. * p < 0.05 vs. sham-operated animals, † p < 0.05 vs. vehicle-treated subtotally nephrectomised rats.

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Fig 1 Expand

Fig 2.

Animal survival.

Kaplan-Meier curves show reduced animal survival in SNX rats when compared with animals that had undergone sham surgery. No effect of dapagliflozin on mortality was noted. * p < 0.05 vs. sham-operated animals.

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Fig 2 Expand

Fig 3.

Glomerulosclerosis.

Representative glomerular images (A-D) and with quantitative analysis (E) of periodic acid-Schiff stained kidney sections. When compared with animals that had undergone sham surgery (A, B), kidney sections from SNX rats (C, D) show substantial glomerulosclerosis that was similar in vehicle (C) and dapagliflozin-treated rats (D). Original magnification x 400. Scale bars: 50 μm. * p < 0.05 vs. sham-operated animals.

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Fig 3 Expand

Fig 4.

Interstitial fibrosis.

Representative cortical tubulointerstitial images (A-D) and with quantitative analysis (E) of Masson’s trichrome-stained kidney sections. When compared with animals that had undergone sham surgery (A, B), kidney sections from SNX rats (C, D) show substantial interstitial fibrosis (blue) that was similar in vehicle (C) and dapagliflozin-treated rats (D). Original magnification x 100. Scale bar: 200 μm. * p < 0.05 vs. sham-operated animals.

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Fig 4 Expand

Fig 5.

Gene expression.

Kidney gene expression of transforming growth factor-ß1, α(I) IV and α(I) I collagen. mRNA was expressed relative to that of RPL13a. The ratio, so-derived was then expressed relative to vehicle-treated rats that had undergone sham surgery that was arbitrarily set at 1. SNX was associated with overexpression of both TGF-ß and α(I) IV collagen that for α(I) I collagen was greater in SNX animals that received dapagliflozin. * p < 0.01 versus sham nephrectomy.

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Fig 5 Expand

Fig 6.

Macrophage infiltration.

Kidney sections from SNX rats revealed abundant ED-1 labelled macrophages, predominantly localized to the interstitium that were more common in SNX rats that had received dapagliflozin. Original magnification x 400. Scale bar: 50 μm. * p < 0.05 vs. sham-operated animals.

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Fig 6 Expand

Fig 7.

Phosphorylated Smad2.

Kidney sections from SNX rats showed abundant nuclear staining for phosphorylated Smad2 in both vehicle and dapagliflozin-treated animals contrasting the dearth of immunopositive cells in sham nephrectomised rats. Original magnification x 160. Scale bar: 100 μm. * p < 0.05 vs. sham-operated animals.

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Fig 7 Expand

Fig 8.

Type I collagen.

Representative sections stained with anti-collagen I antibody revealing minimal immunolablelling in sham rats (A, B) while those from SNX animals (C, D) displayed increased deposition surrounding Bowman’s capsule and in bands of interstitial fibrosis. No difference in immunolabelling was evident between those rats that had or had not received dapagliflozin. Original magnification x 100. Scale bar: 200 μm.

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Fig 8 Expand