Table 1.
Nucleotide sequences of the primers used for real-time polymerase chain reaction.
Fig 1.
A) Renals of UUO rats underwent display significant pathological changes and an increase in collagen I expression. Sprague-Dawley rats were divided into 2 groups (5/group): Sham- operation (SHM) and UUO. All rats were sacrificed at day 14. renal cortex stained with HE, Masson’s trichrome or immunohistochemical for collagen I (×200). Semiquantitative analysis of tubulointerstitial damages as the degree of interstitial collagen deposition and collagen I expression in the renal cortex of rats from each group. Data expressed as means±S.E. *p<0.05 vs. SHM. B) Expression of α-SMA protein was increased and that of E-cadherin protein was decreased in renal cortex of UUO rats. The renal cortexes from different rats were lysed. The cell lysates were blotted with anti-α-SMA and anti-E-cadherin antibody. C) Expression of α-SMA and Collagen I mRNA increased and that of E-cadherin mRNA decreased in renal cortex of UUO rats. The mRNA from different rats was extracted, and the mRNA for α-SMA, E-cadherin, and Collagen I were detected by real-time PCR. Data expressed as means±S.E. *p<0.05 vs. SHM.
Fig 2.
A) Representative images of the cellular localization of GSTA3 in renal cortex of rats (×200), and quantitative analysis of GSTA3 expression. B) The expression of GSTA3 protein and mRNA. C) The cellular localization of GSTA3 in renal cortex of human (×200), and quantitative analysis of GSTA3 expression. Data are expressed as means±S.E of three independent experiments. *p<0.05 vs. Sham.
Table 2.
Differentially expressed proteins in proteomics analysis.
Fig 3.
A) Effects of TGF-β1 on GSTA3, FN, α-SMA, E-cadherin, megalin and p-Smad2/3 protein expressions in NRK-52E cells. Serum-starved NRK-52E cells were stimulated with 10 ng/ml TGF-β1 for 48 h. SB431542 (10ng/ml) was administered 1 h prior to the addition of TGF-β1. The expression of GSTA3, FN, α-SMA, and E-cadherin was subjected to western blot analysis. B) Effects of TGF-β1 on GSTA3, FN, α-SMA, E-cadherin and megalin mRNA expressions in NRK-52E cells. The mRNA from NRK-52E cells was extracted, and the mRNA expression for GSTA3, α-SMA, and E-cadherin was detected by real-time PCR.
Fig 4.
A) Effects of knocking down GSTA3 on EMT in NRK-52E cells. After a 6 h incubation of cells with siRNA-Lipofectamine 2000 complex, the cells were maintained with normal DMEM for an additional 18 h. Cells were treated with TGF-β1 and harvested for GSTA3, FN, E-cadherin, α-SMA and megalin protein expression after 48 h. B) Effects of modulation of GSTA3 over-expression on EMT in TGF-β1 treated NRK-52E cells. After the transfected cells were treated with 10 ng/ml TGF-β1 for 48 h, GSTA3, FN, E-cadherin, α-SMA, megalin and p-Smad2/3 protein expression were analyzed by western blot. Results are presented as the means±S.E. of three independent experiments. *p<0.05 vs. control; #p<0.05 vs. TGF-β1.
Fig 5.
Effect of GSTA3 on NRK-52E cellular ROS.
A) ROS level of knocking down GSTA3 in NRK-52E; B) ROS level of treatment with 10 ng/ml TGF-β1 for 15min in GSTA3 over-expression cells. C) SOD activity knocking down GSTA3 in NRK-52E; D) SOD activity of treatment with 10 ng/ml TGF-β1 for 15min in GSTA3 over-expression cells. Results are presented as the means±S.E. of 3 independent experiments. *p<0.05 vs. control; #p<0.05 vs. TGF-β1.