Figure 4.

The effect of CYD-6-17 on PDPK1 expression and its role in modulating β-catenin expression. (a) The expression of PDPK1 mRNA and protein in RCC cells treated with CYD-6-17. (b) PDPK1 gene promoter activity determined by luciferase reporter assay in RCC cells treated with CYD-6-17. (c) Cells were transfected with different dose of PDPK1 cDNA for 24 h and then treated with CYD-6-17 (0.5 μM) for 48 h. MTT assays were performed to determine cell viability. (d) The mechanism of PDPK1 in modulating GSK3β activity. HK-2 KD and Sor001 cells were transiently transfected with 2.5 μg of vector control, WT, or KI PDPK1 for 24 h, western blot analyses were performed to determine the expression of PDPK1, total GSK3β or phosphorylated GSK3β, and β-catenin levels (left panel). The relative cell number treated with CYD-6-17 (0.5 μM) for 48 h was determined by MTT assay (right panel). *P<0.05