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. 2011 Oct 24;6(10):e26615. doi: 10.1371/journal.pone.0026615

Figure 2. Phenethyl isothiocyanate (PEITC) treatment increases transcriptional activity of Notch in malignant and normal prostate cells.

Figure 2

(A) Effect of PEITC treatment on RBP-Jk luciferase reporter activity (a measure of transcriptional activity of Notch) in LNCaP, PC-3, DU145, and PrEC cells after 8- or 16-hour treatment with dimethyl sulfoxide (DMSO) or 5 µM PEITC. Results shown are mean ± SD (n = 3). *Significantly different (P<0.05) compared with DMSO-treated control by one-way ANOVA followed by Dunnett's test (LNCaP and PC-3) or Student's t-test (DU145 and PrEC). (B) Immunoblotting for cleaved Notch1 and cleaved Notch2 using lysates from DU145 and PrEC cells treated with DMSO (control) or PEITC (2.5 or 5 µM) for the indicated time periods. Blots were stripped and re-probed with anti-actin antibody. Arrow in panel B (DU145 cells) identifies cleaved Notch2, the lower band is non-specific based on siRNA results shown in Fig. 4A. Immunoblotting for each protein was done at least twice using independently prepared lysates. Numbers above bands represent changes in protein levels relative to corresponding DMSO-treated control. Each experiment was repeated at least twice.