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. 2011 Aug 18;17(11-12):1275–1284. doi: 10.2119/molmed.2011.00125

Figure 7.

Figure 7

Interactions between Ahr and Wt1 during the course of renal cell differentiation. (A) Downregulation of markers of differentiation by Wt1 siRNA. mK4 cells were cultured in the presence of Wt1 siRNA for 3 d. qRT-PCR values were calculated to represent fold change normalized to 18S compared with nonspecific siRNA control. sfrp-1 and lim 1 homeobox gene were used as markers of mesenchymal identity, while Igf-1 rec, Igf-2 rec, Wnt4, and E-cadherin were used as markers of epithelial identity. (B) Deregulation of Wt1 targets by Wt1 siRNA. mK4 cells were cultured in the presence of Wt1 siRNA for 3 d. qRT-PCR values were calculated to represent fold change normalized to 18S compared with nonspecific siRNA control. Syndecan1, paired box protein 2, EGF rec, retinoic acid receptor alpha, taurine transporter, and Wilms tumor transcription factor. (C) The mK4 cells were transfected for 24 h with the CB6-Wt1 (−) exon 5/(−) exon 9 plasmid encoding for 52 kDa Wt1 protein or empty vector after transfection with Ahr siRNA. Reduced Wt1 protein levels following Ahr knockdown were rescued in cells transfected with Wt1 cDNA compared with empty vector. (D) Real time PCR shows decreased mRNA levels for Igf-1 rec. and Wnt-4 following Ahr knockdown and effective rescue in cells transfected with Wt1 cDNA; however, decreases in sFrp-1 mRNA following Ahr knockdown were not restored in cells transfected with Wt1 cDNA. One representative experiment out of three is shown. NT = no treatment.