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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: Interplay between intergrin-linked kinase and ribonuclease inhibitor affects growth and metastasis of bladder cancer through signaling ILK pathways

Fig. 5

Up-regulating RI and ILK regulates the expression of ILK signaling pathway molecules and proteins associated with EMT in vitro. The total protein of cells was collected using cell lysis buffer, the expressions of ILK signaling pathway molecules and proteins associated with EMT were examined using corresponding antibodies by immunoblot analysis. β-actin was served as the control. Relative protein levels were normalized against those of β-actin by MJ Opticon Monitor Analysis Software (Bio-Rad). Data were calculated as means ± S.D (n = 3). *P < 0.05. a & b The results demonstrated that transfections with pCMV-3 × FLAG-ILK in EJ cells significantly up-regulated the expressions of p-Akt (S473), p-GSK3β (S9), p-PI3K, p-PTEN, p-mTOR and β-catenin, compared with control groups respectively, whereas these protein levels decreased obviously in EJ-RI cells. The protein expressions of AKT, PI3K, mTOR and GSK3β did not change apparently, compared with control groups respectively. c & d Western blot assay and semi-quantitative analysis showed that protein levels of MMP-2, MMP-9, N-cadherin, Snail, Twist and Vimentin were decreased, while protein levels of E-cadherin and Smad2 were significantly increased in EJ-RI cells compared with the control groups respectively. E-cadherin was significantly weaker in the EJ-ILK cells group compared with the other control groups and MMP-2, MMP-9, N-cadherin, Snail, Twist, S100A4 and Vimentin were increased in EJ-ILK cells groups

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