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. 2014 Nov 20;16(11):939–949. doi: 10.1016/j.neo.2014.09.005

Figure 2.

Figure 2

Endogenous galectin-3 inhibits ALP expression. (A) Cancer-secreted galectin-3 accumulated in condition medium in a time-dependent manner. PC-3 cells were cultured with condition medium for 0 to 2 days, and then it was subjected to immunoblot and ELISA without concentration. (B and C) Secretory factor by PC-3 cells inhibits ALP expression and its enzymatic activity in a sugar-dependent manner. Condition medium from PC-3 cell culture was concentrated to 20-fold, and 5 μl of concentrated medium was added into osteoblast cell culture in a 96-well plate. Culture medium was changed every 3 days. After 1 week, it was subjected to ALP stain and pNPP assay. The suppressive effect of cancer-secretory factor on osteoblast differentiation was recovered by the treatment of lactose (75 mM). (D and E) Inhibitory effect of secretory factors from PC-3 cells on osteoblast differentiation is mediated by galectin-3. Osteoblast cells were subjected to ALP stain and pNPP assay after co-culture with PC-3 cells for 1 week. MCP, a specific inhibitor of galectin-3, recovered the downregulated ALP expression and its activity. Data represent means ± SD. *P < .05 versus vehicle control.