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. 2012;7(10):e46442.
doi: 10.1371/journal.pone.0046442. Epub 2012 Oct 3.

Renalase's expression and distribution in renal tissue and cells

Feng Wang  1 , Tao Xing, Junhui Li, Mei Bai, Ruimin Hu, Zhonghua Zhao, Shoufu Tian, Zhigang Zhang, Niansong Wang
Affiliations

Renalase's expression and distribution in renal tissue and cells

Feng Wang et al. PLoS One. 2012.

Abstract

To study renalase's expression and distribution in renal tissues and cells, renalase coded DNA vaccine was constructed, and anti-renalase monoclonal antibodies were produced using DNA immunization and hybridoma technique, followed by further investigation with immunological testing and western blotting to detect the expression and distribution of renalase among the renal tissue and cells. Anti-renalase monoclonal antibodies were successfully prepared by using DNA immunization technique. Further studies with anti-renalase monoclonal antibody showed that renalase expressed in glomeruli, tubule, mesangial cells, podocytes, renal tubule epithelial cells and its cells supernatant. Renalase is wildly expressed in kidney, including glomeruli, tubule, mesangial cells, podocytes and tubule epithelial cells, and may be secreted by tubule epithelial cells primarily.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Western blotting of the expression of pBudCE4.1-Renalase in 293T cells.
1: empty pBudCE4.1vector, 2: pBudCE4.1-Renalase.
Figure 2
Figure 2. Western blotting verification of anti-renalase monoclonal antibody.
1: recombinant renalase protein.
Figure 3
Figure 3. Immunofluorescence testing of renalase's expression in renal tissue.
A: renalase expressed in glomeruli (Anti-renalase monoclonal antibody was used as primary antibody) (400×). B: glomeruli negative control (PBS was used as control) (400×). C: renalase expressed in renal proximal tubules (Anti-renalase monoclonal antibody was used as primary antibody) (400×). D: renal proximal tubule negative control (PBS was used as control) (400×).
Figure 4
Figure 4. Immunohistochemical detection of renalase's expression in kidney tissues.
A: renalase expressed in glomeruli (Anti-renalase monoclonal antibody was used as primary antibody) (400×). B: glomeruli negative control (PBS was used as control) (400×). C: renalase expressed in renal proximal tubules (Anti-renalase monoclonal antibody was used as primary antibody) (400×). D: Renal proximal tubule negative control (PBS was used as control) (400×).
Figure 5
Figure 5. Immunofluorescence testing renalase's expression in kidney cells.
A: renalase expressed in mesangial cells (Anti-renalase monoclonal antibody was used as primary antibody) (400×). B: mesangial cells negative control (PBS was used as control) (400×). C: renalase expressed in podocytes (anti-renalase monoclonal antibody was used as primary antibody) (400×). D: podocytes negative control (PBS was used as control) (400×). E: renalase expressed in renal tubular epithelial cells (anti-renalase monoclonal antibody was used as primary antibody) (400×). F: renal tubular epithelial cells negative control (PBS was used as control) (400×).
Figure 6
Figure 6. Western blotting testing renalase's expression in kidney cells.
Anti-renalase monoclonal antibody was used as primary antibody. A: mesangial cells. B: culture supernatant of mesangial cells. C: podocytes. D: cuture supernatant of podocytes. E: renal tubular epithelial cells. F: culture supernatant of renal tubular epithelial cells.
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