Fig. 3.

Direct co-culture of PCa cells with MSCs significantly up-regulated expression of Notch signaling-related genes in PCa cells. Two different PCa cell lines (PC3, left; LNCaP, right) were employed to establish the mono-culture, transwell-culture, and mixed co-culture with MSCs system. Cells were collected after treatment for 48 h. a mRNA levels of Notch targets (Hes1 and Hey1) were determined by quantitative RT-PCR (q-PCR). b, c mRNA and protein expression levels of Notch receptor (Notch1) expressed in PCa cells were detected by qRT-PCR and western blot analyses. d NICD level was detected by western blotting in PC-3 and LNCaP cells, with β-actin as a loading control. e, f mRNA and protein expression levels of Notch1 ligand (Jagged 1) expressed in MSCs were detected by qRT-PCR and western blot analyses. Data are presented as mean ± SD; *P < 0.05, **P < 0.01; ***P < 0.001