Figure 4.

LEF-1-ΔL reduced the abilities of colony formation and adhesion in SW480 and HT29 cells. (A) and (C) Colony-forming assay. Fifty SW480 and HT29 cells expressing LEF-1-ΔL or LEF-1-FL were cultured in 35-mm plates, and colonies with more than 50 cells were counted on day 14 after the start of the culture. (B) and (D) Colonies in A and C were counted under light microscopy. Data are expressed as mean ± S.D. *P < 0.01. (E) and (F) 2 × 10⁴ cells in A and C were added to wells in a 96-well plate, which was coated with BSA and matrigel. After incubation in a CO₂ incubator at 37°C for 1 h, non-adherent cells were removed by washing with PBS. The number of adherent cells was counted with the MTT assay. (G) and (H) Cell migration assay. SW480 and HT29 cells expressing LEF-1-ΔL or LEF-1-FL were seeded in the upper chamber of a trans-well culture system, with SDF1a (100 ng/ml) in the lower chamber. Colon cells migrating into the lower chamber were analyzed by FACS and cell counting. (I) and (J) The expression of CXCR4 in cells in A and B was analyzed by FACS.