TAK1 inhibition limits the expression of mesenchymal markers by MCs derived from peritoneal effluent of PD patients.

<p><b>A</b>, Confocal immunofluorescence (red) of MCs from a patient undergoing PD. Cells were treated with DMSO or NP-009245 (600 nM) for 72 h. Cells were fixed and stained with a monoclonal antibody against fibronectin. Nuclei were stained with Hoechst 33342 (blue). The immunofluorescence shown is representative of three independent experiments. <b>B</b>, Western blots showing the expression of fibronectin, E-cadherin and vimentin in total cell lysates of MCs treated with DMSO, NP-009245 (600 nM) or CI-1040 (2 µM) for 72 h. Expression of tubulin was used as a loading control. The results shown are representative of three independent experiments. <b>C</b>, Effect of TAK1 pharmacological inhibition on fibronectin and E-cadherin mRNA expression in MCs from patients undergoing PD. Quantitative RT-PCR was performed on total RNA from MCs treated as in A. Histone H3 mRNA levels were used for normalization. Bars represent means+s. e. m. of duplicate determinations in three independent experiments. <b>D</b>, Western blots showing the expression of E-cadherin, fibronectin and TAK1 in total cell lysates of MCs transfected with either control or specific TAK1-targeting siRNAs. Scr, cells transfected with control siRNA. Data are representative of three independent experiments. <b>E</b>, Effect of TAK1 siRNA silencing on E-cadherin mRNA expression in MCs. Cells were treated as in D. Quantitative RT-PCR was performed; bars represent means+s. e. m. of duplicate determinations in three independent experiments. * p<0.05 versus siScrambled treated cells.