Figure 2.

RASAL2 suppresses stemness in BCa cells. (a) Quantitative real-time RT-PCR and western blotting analysis of RASAL2 levels in human BCa cell lines. (b) Quantitative real-time RT-PCR and western blotting analysis of RASAL2 levels in 5637 cells transfected with RASAL2 shRNAs and scramble shRNA, or T24 cells infected with RASAL2 lentivirus and scramble control. (c) Tumorsphere assay of 5637 cells transfected with RASAL2 shRNAs and scramble shRNA, or T24 cells infected with RASAL2 lentivirus and scramble control. The tumorsphere number was counted and plotted, and percentage of tumorspheres with diameters <50 μm, 50–100 μm or >100 μm was calculated and plotted. The scale bar represents 100 μm. (d) Colony formation assay of 5637 cells transfected with RASAL2 shRNAs and scramble shRNA, or T24 cells infected with RASAL2 lentivirus and scramble control. The colonies number was counted and plotted. (e) Western blotting analysis of RASAL2, CD44 and SOX2 in 5637 cells transfected with RASAL2 shRNAs and scramble shRNA, or T24 cells infected with RASAL2 lentivirus and scramble control. GAPDH was used as internal loading control. *P<0.05, **P<0.01