Figure 1.

OCCM30 cultured on fibrin enhances canonical Wnt signaling. OCCM30 cells were cultured on fibrin and tissue culture dish (TCD) in differentiation media over a three-day period with/without LiCl. Fibrin induced the translocation of β-catenin from the cytoplasm to the nucleus in cementoblasts (green-fluorescence represented β-catenin): OCCM-30 cells were cultured on fibrin and TCD for three days, and the nuclear translocation of β-catenin detected by immunofluorescence showed that the protein translocated to the nucleus was increased in the cells cultured on fibrin (A). β-catenin expression was also determined by immunoblot analysis and it was highly expressed rather than TCD (B). Cementoblast cells were transfected with Top flash (wild-type promoter) or Fop flash (mutant promoter) reporter plasmids and cultured in differentiation media for the last 24 h, and luciferase activities were measured in cell lysates and normalized to a Renilla transfection control (C). Fibrin exhibited a higher expression of Wnt3a and induced canonical Wnt signaling which was determined by quantitative real-time polymerase chain reaction (PCR) and western blot analysis (D). Each experiment was repeated two times independently. * Significantly different (p < 0.05).