3.6 ISX9 upregulates the expression of Wnt target genes Axin2, LEF1, Fibronectin, and Survivin
Considering that activation of Wnt/β-catenin signaling will upregulate the transcription of Wnt target genes (Mosimann, Hausmann, & Basler, 2009), we assessed the effect of ISX9 on mRNA expression of Wnt target genes Axin2, LEF1, Fibronectin, and Survivin. HEK293T, HaCat and NIH3T3 cells were treated with increasing concentrations of ISX9 (2.5-40 µM) for 24 h. Real-time PCR analysis showed that ISX9 dose-dependently enhanced mRNA expression of Axin2, LEF1, Fibronectin, and Survivin in all three cell lines (Figure 5A-C). Consistent with mRNA data, we detected increased protein levels of Axin2, LEF1, and Survivin in response to ISX9 treatment in all three cell lines (Figure 5D-F). These results confirmed the activation of Wnt/β-catenin signaling in HEK293T, HaCat and NIH3T3 cells treated with ISX9.