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Disruption of tissue-type plasminogen activator gene in mice reduces renal interstitial fibrosis in obstructive nephropathy
Junwei Yang, … , Kari Nejak, Youhua Liu
Junwei Yang, … , Kari Nejak, Youhua Liu
Published November 15, 2002
Citation Information: J Clin Invest. 2002;110(10):1525-1538. https://doi.org/10.1172/JCI16219.
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Article Nephrology

Disruption of tissue-type plasminogen activator gene in mice reduces renal interstitial fibrosis in obstructive nephropathy

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Abstract

Research Article

Authors

Junwei Yang, Ryan W. Shultz, Wendy M. Mars, Rodney E. Wegner, Yingjian Li, Chunsun Dai, Kari Nejak, Youhua Liu

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Figure 10

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tPA induces MMP-9 protein secretion and gene expression in renal interst...
tPA induces MMP-9 protein secretion and gene expression in renal interstitial fibroblasts. (a and b) Zymographic analysis shows an induction of MMP-9 secretion by tPA. Rat renal interstitial fibroblasts (NRK-49F) were incubated with different concentrations (10–9 to 10–7 M) of human tPA protein for 48 hours (a), or with 10–8 M tPA for various periods of time (b). Samples of the cell culture supernatants were separated on a polyacrylamide gel containing 1 mg/ml gelatin. Proteolytic activity was demonstrated by digestion of gelatin, resulting in the bands of clearing. The locations of bands corresponding to MMP-9 and MMP-2 are indicated. (c and d) RT-PCR analysis demonstrates that tPA induces MMP-9 gene expression in renal interstitial fibroblasts. NRK-49F cells were incubated with different concentrations (10–9 and 10–8 M) of human tPA protein for 24 hours (c), or with 10–8 M tPA for various periods of time (d). Cellular MMP-9 mRNA levels were determined by RT-PCR analysis. Amplified MMP-9 cDNA fragment with the correct size (1.16 kb) is indicated. RT-PCR amplification of housekeeping β-actin was performed in an identical manner to serve as control. (e and f) Southern blot analysis of the RT-PCR product. To confirm the RT-PCR product as MMP-9 cDNA, Southern blot analysis of the RT-PCR product in c and d was performed using an internal oligonucleotide corresponding to MMP-9 cDNA as a probe.

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ISSN: 0021-9738 (print), 1558-8238 (online)

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