Fig. 4
Regulation of ALPL and ANKH expression by bFGF in SHEDs. Cells were maintained in normal growth medium in the presence or absence of 10 ng/mL bFGF for 6, 12, or 24 h. a Addition of bFGF significantly inhibits ALPL mRNA by 24 h. b The induction of ANKH mRNA by bFGF is observed at 6, 12, and 24 h. c, d For both ALPL and ANKH, inclusion of 20 mM FGFR inhibitor (SU5402) in growth medium abolishes effects of bFGF at 24 h. Addition of 1 μg/ml cyclohexamide (CHX) inhibits effects of bFGF on ALPL and ANKH expression, suggesting that protein translation is required. e In culture medium collected from SHEDs on days 1, 6, and 14 after osteogenic induction, bFGF significantly reduces Pi/PPi ratio by day 14. f Addition of 0.05 U ALP rescues bFGF-attenuated ALP expression and mineral deposition in SHEDs. The Mann-Whitney U test was employed for comparisons of two groups. The Kruskal-Wallis test followed by pairwise comparison was used for comparisons of three or more groups. Bars indicate statistically significant differences (*p < 0.05, **p < 0.01). OM osteogenic induction medium