Fig. 4

Inhibition of the Notch pathway inhibited BMSCs-induced stemness characteristics of PCa cells through cell–cell contact. Two different PCa cell lines (PC3, a–d; LNCaP, e, f) were employed to establish the mono-culture, transwell-culture, and mixed co-culture with the MSCs system. After treatment with the inhibitor of Notch1 LY3039478 (100 nM) for 72 h, the PCa cells were collected from each group and subjected to flow cytometry a, colony formation b, sphere formation assay c and limited dilution of clone formation d. The collected LNCaP cells were subjected to colony formation e and sphere formation assay f. g, h PC-3 cells were employed to establish the mono-culture, transwell-culture, and mixed co-culture with the MSCs system. After treatment with IMR-1 (15 uM) for 72 h, the PC-3 cells were collected from each group and then colony formation g, and sphere formation assays h were carried out. Data are presented as the mean ± SD; *P < 0.05, **P < 0.01; ***P < 0.001