Fig. 6

PLCε regulates Twist1 through nuclear translocation of PPARβ. a PC3 cells were transfected with DDK-Twist1 plasmid. After 12 h, cells were treated with 100 μM cycloheximide (CHX) for time periods as indicated. Western blotting analysis was conducted with DDK and β-actin antibodies. Densitometric values were determined and presented. The half-life (50%) of DDK-Twist1 was indicated. b PC3 cells were infected with sh-PLCε lentiviral and treated with 25 μM MG-132 for 24 h, and cell lysates were collected for western blotting and protein quantification analyses. c, d Western blotting and protein quantification analyses detected the expression of PLCε, PPARβ, PPARα and PPARγ in sh-PLCε-infected PC3 cells. e, f The protein and mRNA expression of PLCε and PPARβ after knockdown of PPARβ in PC3 cells line. g, h Western blotting and protein quantification analyses detected the expression of PPARβ and Twist1 in sh-PLCε-infected PC3 cells. i Twist1 promoter transcriptional activity determined by luciferase assay in PC3 cells. j Immunofluorescence staining demonstrated PPARβ intracellular distribution in three PCa cell lines.Magnification × 400. Bars = 50 μm. k A nuclear-cytoplasmic separation was performed in PCa cells. Western blotting analysis of each fraction detected the expression of PPARβ. β-actin and Histone (H) 3 were used as an internal control. Data were represented as mean ± SD. of three individual experiments. *p < 0.05, **p < 0.01, and *** p < 0.001 vs. Controls