Fig. 5.

Removal of HMW-HA from estrogen-deficient culture medium increased the RANKL expression in ST2 cells. A Experimental plan. Three hundred thousand cells were incubated for 12 h in RPMI-1640 medium. After cells were attached to the dishes, culture media were replaced with (I) charcoal-stripped FBS and (II) charcoal-stripped FBS with 100 nM of hyaluronidase for 12 h or 24 h. B, C Relative expression of RANKL and OPG in the presence or absence of hyaluronidase at 12 and 24 h. Colorful lines are associated with each 12 and 24 h samples. D Average ratio of RANKL mRNA to OPG mRNA in the presence or absence of hyaluronidase. Data are expressed as mean ± SD, analyzed by Wilcoxon signed-rank test (B, C), Wilcoxon rank-sum test (D), showing all data points. *p < 0.05. ns, not significant. n = 6 per group. Numerical data are presented in Supplementary Table 6