Galectin-3 Plays an Important Role in BMP7-Induced Cementoblastic Differentiation of Human Periodontal Ligament Cells by Interacting with Extracellular Components

Figure 4

Galectin-3/CM3 was accumulated in cell membrane fraction during BMP7-induced cementoblastic differentiation. For induction of cementoblastic differentiation, hPDLCs were treated with 100 ng/mL BMP7 and 10 μM SB431542 for 9 days. Cells were harvested at the indicated times, and cytosolic and membrane fractions were prepared as mentioned in Materials and Methods. (a) Galectin-3 was detected in both cytosolic and membrane fractions in mouse cementoblasts (A) and in BMP7-induced human cementoblast-like cells (B). total: total extract; cyto: cytosolic fraction; mem: membrane fraction. In (B), BMP7 and SB431542 (B7/SB) were treated in hPDLCs for 0 day (lanes 1, 4, and 7), 4 days (lanes 2, 5, and 8), and 9 days (lanes 3, 6, and 9). Integrin β1 (ITGβ1) was used as a marker for membrane fraction in western blots. (b) Subcellular localization of galectin-3/CM3 in BMP7-induced human cementoblast-like cells. (A) Immunostaining with Cy3-galectin-3 and FITC-calnexin; (B) immunostaining with Cy3-galectin-3 and FITC-GM130; (C) immunostaining with Cy3-galectin-3 and FITC-phalloidin.

(a)

(b)