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Real time PCR and Western blot verification in SC cultures of altered expression of cell polarity and Wnt signaling genes following Wt1 ablation.

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posted on 2013-08-01, 04:02 authored by Xiao Na Wang, Ze Song Li, Yu Ren, Tao Jiang, Ya Qing Wang, Min Chen, Jun Zhang, Jian Xiu Hao, Yan Bo Wang, Ri Na Sha, Yi Huang, Xiao Liu, Jing Chu Hu, Guang Qing Sun, Hong Gang Li, Cheng Liang Xiong, Jun Xie, Zhi Mao Jiang, Zhi Ming Cai, Jun Wang, Jian Wang, Vicki Huff, Yao Ting Gui, Fei Gao

(A) The expression of cell polarity related genes (E-cadherin, Par6b, Cdc42bp5, Sfn) and Wnt signaling genes (Wnt4 and Wnt11) is significantly decreased in Wt1-deficient SCs (white bars). (B) E-cadherin and Espin protein expression is significantly reduced in Wt1-deficient SCs, whereas the expression of occludin, N-cadherin, Zo-1, and β-catenin is not changed. (C) The expression of E-cadherin, Par6b, Wnt4, Wnt11, and Cdc42ep5 in Wt1-deficient SCs was rescued by transfection with Wt1 expressing adenovirus. (D) The expression of Par6b was induced by Wnt4 and Wnt11 expressing adenovirus in Wt1-deficient SCs. (E) The expression of E-cadherin was not rescued by Wnt4 and Wnt11 expressing adenovirus in Wt1-deficient SCs. (F) Wnt4 was knocked down by two different siRNAs (siWnt4-2 and siWnt4-3) in SCs and the expression of E-cadherin and Par6b was significantly decreased compared to control cells (NC) treated with a scrambled siRNA. * p<0.05.

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