1. Volume 75, Issue 12, Pages 1288-1296 (June 2009)
Overexpression of cytochrome P450 4F2 in mice increases 20-hydroxyeicosatetraenoic acid production and arterial blood pressure 
Xiaoliang Liu, Yanyan Zhao, Luzeng Wang, Xianghong Yang, Zhihong Zheng, Yuanyuan Zhang, Fangjie Chen, Hong Liu 
Kidney International 
Volume 75, Issue 12, Pages (June 2009)
DOI: /ki
Copyright © 2009 International Society of Nephrology Terms and Conditions

2. Figure 1
KAP promoter activity in transfected HEK293 cells. (a) Relative luciferase activity assay. Using the Dual Luciferase Reporter assay system, the relative luciferase activity is presented as the ratio of luciferase activity of firefly to that of Renilla as internal control. (b) Western blot analysis of CYP4F2 expression. CYP4F2 level was determined by anti-CYP4F2 antibody and anti-his antibody in cells transfected with the pKAP-CYP4F2/his (+) and the pGL3-enhancer (-). The 53-kD band represented CYP4F2 expression, and the 36-kD GAPDH was shown as internal control. Data are presented as mean±s.d. All experiments were performed three times independently.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

3. Figure 2
Schematic map of the linear pKAP-CYP4F2/his and Southern blot analysis of CYP4F2 in transgenic mice. (a) The fragment for microinjection contained the 224-bp KAP promoter, CYP4F2 cDNA, 6 × his tag, and SV40 enhancer. Arrow-indicated P1 and P2 are the primers specific for CYP4F2 to screen positive mice by PCR. (b) Southern blot analysis. The hybridization bands of tail DNA with specific CYP4F2 probe confirmed the integration of the transgene in the three transgenic lines. WT: wild-type.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

4. Figure 3
CYP4F2 expression by Western blot analysis. 53-kD CYP4F2 was detected using anti-his antibody, and 36-kD GAPDH is shown as an internal control. (a) CYP4F2 expression in the kidney of three transgenic lines and wild-type (WT) control. (b) CYP4F2 expression profile of female offspring in line F0-16. (c) CYP4F2 expression profile of male offspring in line F0-16. (d) Scanning densitometry of b and c. Data are presented as mean±s.d. All experiments were performed three times independently.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

5. Figure 4
CYP4F2 expression by immunohistochemistry. (b, d, f and h) Kidney, liver, brain, and uterus of transgenic (TG) mice, and (a, c, e and g) wild-type (WT) controls, respectively. Anti-his antibody was used as primary antibody and positive stainings are indicated by arrows, using Diaminobenzidin as the chromagen. Micrographs were taken at × 200 magnification. The bar represents 100 μm. (1) glomerulus; (2) proximal tubule; (3) lobular central vein; (4) hepatocyte of the lobule; (5) endometrium.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

6. Figure 5
The ω-hydroxylation analysis of renal microsomes. (a) Increased 20-HETE production by transgenic renal microsomes. In the AA ω-hydroxylation reactions, the 20-HETE production was significantly higher in the transgenic (TG) group (660 (408–1073)) than in the wild-type (WT) group (298 (224–398)). (b) CYP4F2 expression in renal microsomes by Western blot analysis. The 53-kD band of CYP4F2 was detected by anti-CYP4F2 antibody and anti-his antibody, and the 36-kD GAPDH was shown as an internal control. All experiments were performed three times independently.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

7. Figure 6
SBP of CYP4F2 transgenic mice. (a) Comparison of SBP in CYP4F2 transgenic mice. The SBPs of transgenic mice from the three transgenic lines were compared with the wild-type (WT) controls. Data are presented as mean±s.d. *P<0.001 in the analysis of variance. (b) Correlation of logarithmic urinary 20-HETE with SBP. Pearson's correlation coefficient and P-value are indicated on the graph.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

8. Figure 7
Time course blood pressure measurement. The blood pressure of transgenic (TG) mice in line F0-16 (n=12) and wild-type (WT) controls (n=9) aged 8 to 40 weeks was measured at four-week intervals to record the development of hypertension. The SBP of TG mice was elevated in early life and maintained at higher levels than WT controls throughout late life.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions