Table 1.
Mouse specific primers used in quantitative real-time PCR.
Fig 1.
Maternal body weight at the commencement of breeding.
Pre-breeding body weight of mothers. Results expressed as mean ± SEM, n = 13–20. ****p<0.0001.
Table 2.
Biometric parameters of week 9 offspring.
Table 3.
Biometric parameters of week 32 offspring.
Table 4.
Metabolic parameters of week 9 offspring.
Table 5.
Metabolic parameters of week 32 offspring.
Fig 2.
(A) Renal global DNA methylation expressed as % 5mC of total DNA in week 9 offspring, (B-D) week 9 offspring mRNA expression of DNMT1, DNMT3a and Tet3, (E) renal global DNA methylation expressed as % 5mC of total DNA in week 32 offspring, (F-H) week 32 offspring mRNA expression of DNMT1, DNMT3a and Tet3. Results expressed as mean ± SEM, n = 4–6. *p<0.05, ***p<0.001.
Fig 3.
(A, B) Week 9 offspring representative images of PAS staining for glomerulosclerosis at 400x magnification, and glomerulosclerosis index, (C, D) week 32 offspring representative images of PAS staining for glomerulosclerosis at 400x magnification, and glomerulosclerosis index. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 4.
Renal fibrosis as measured by Picrosirius Red (PSR) staining.
(A) Week 9 offspring representative images of PSR staining at 200x magnification, and quantitation, (B) week 32 offspring representative images of PSR staining at 200x magnification, and quantitation. Results expressed as mean ± SEM, n = 5–6, **p<0.01.
Fig 5.
Kidneys assessed by PAS and Picrosirius Red staining. (A, B) Tubulointerstitial fibrosis assessment at week 9 and week 32, (C, D) tubular vacuolation assessment at week 9 and week 32, (E, F) tubular dilatation assessment at week 9 and week 32, (G, H) casts scoring at week 9 and week 32. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 6.
Renal fibrotic changes as measured by collagen III.
(A) mRNA expression of collagen III at week 9, (B) western blotting for collagen III at week 9, (C) mRNA expression of collagen III at week 32, (D) western blotting for collagen III at week 32. Results expressed as mean ± SEM, n = 6. *p<0.05, ***p<0.001, ****p<0.0001.
Fig 7.
Renal fibrotic changes as measured by collagen III immunohistochemistry.
(A) Week 9 offspring representative images of collagen III staining at 200x magnification, and quantitation (B) Week 32 offspring representative images of collagen III staining at 200x magnification, and quantitation. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 8.
Renal fibrotic changes as measured by collagen IV.
(A) Week 9 offspring mRNA expression of collagen IV, (B) week 9 offspring western blotting for collagen IV, (C) week 32 offspring mRNA expression of collagen IV, (D) week 32 offspring western blotting for collagen IV. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 9.
Renal fibrotic changes as measured by collagen IV immunohistochemistry.
(A) Week 9 offspring representative images of collagen IV staining at 200x magnification, and quantitation (B) week 32 offspring representative images of collagen IV staining at 200x magnification, and quantitation. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 10.
Renal fibrotic changes as measured by fibronectin (FN).
(A) Week 9 offspring mRNA expression of FN, (B) week 9 offspring western blotting for FN, (C) week 32 offspring mRNA expression of FN, (D) week 32 offspring western blotting for FN. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 11.
Renal fibrotic changes as measured by fibronectin (FN) immunohistochemistry.
(A) Week 9 offspring representative images of FN staining at 200x magnification, and quantitation (B) week 32 offspring representative images of FN staining at 200x magnification, and quantitation. Results expressed as mean ± SEM, n = 6, *p<0.05.
Fig 12.
Markers of renal oxidative stress.
(A) Nox2 mRNA expression at week 9, (B) catalase mRNA expression at week 9, (C) MnSOD expression measured by western blotting at week 9, (D) immunohistochemistry for nitrotyrosine at week 9, (E) immunohistochemistry for 8-OHdG at week 9, (F) Nox2 mRNA expression at week 32, (G) catalase mRNA expression at week 32, (H) MnSOD expression measured by western blotting at week 32, (I) immunohistochemistry for nitrotyrosine at week 32, (J) immunohistochemistry for 8-OHdG at week 32. Results expressed as mean ± SEM, n = 6. *p<0.05.
Fig 13.
(A, B) Renal MCP-1 mRNA expression in offspring at week 9 and week 32, (C, D) Renal CD68 mRNA expression in offspring at week 9 and week 32. Results expressed as mean ± SEM, n = 6.