Fig. 4

Characterization of stemness phenotype of cancer cell–derived spheres formed on agar-coated dishes. a and b Reverse transcription quantitative polymerase chain reaction analysis of prostate cancer stem-like cell (PCSC)-associated markers expressed in non-adherent three-dimensional cultured spheres and their corresponding two-dimensional (2D) cultured adherent cells. Results showed that the LNCaP/DU145-derived prostatospheres expressed significantly higher levels of PCSC-associated markers but lower levels of prostate-specific differentiation markers (AR, KLK3/PSA, and KLK2) as compared with their corresponding 2D culture adherent cells. c and d Fluorescence-activated cell sorting analysis of prostatospheres for CD133 and CD44 expressions. c Polychromatic plots and d proportion of cell population graphs of CD133⁺/CD44⁺ cells. Results showed that there was a significant increase of CD133⁺/CD44⁺ cell populations in both LNCaP and DU145 prostatospheres as compared with their corresponding 2D culture adherent cells. *P <0.05, compared with corresponding adherent cells.