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. 2016 Apr 22;17(4):613. doi: 10.3390/ijms17040613

Figure 3.

Figure 3

Heparin has no effect on the production of RANKL, OPG, and M-CSF. (A1–A3) The concentration of factors in the supernatant of the co-culture system, determined with ELISA at 48 h. The medium concentrations of RANKL, OPG, and M-CSF in group 0 were 416, 164, and 1327 pg/mL. After normalized by total proteins, no significant difference among each group (p > 0.05); (B1–B3) The relative expression of RANKL, OPG, and M-CSF mRNA in co-culture cells. The values are expressed as relative expression levels compared with the 0 h groups. At each time point, no significant difference between group 0 and group 10 (p > 0.05); (C1–C3) concentration of the factors in the supernatant of the co-culture system at various time point, determined with ELISA. At each time point, no significant difference between group 0 and group 10 (p > 0.05); and (D) MLO-Y4 and RAW 264.7 were co-cultured with or without 10 IU/mL heparin for eight days. The protein expression of RANKL and M-CSF were analyzed by Western blot at day 0, 2, 4, and 8. Relative band intensity of RANKL and OPG to β-actin were quantified and no significant difference between group 0 and group 10 at each time point (n = 3; p > 0.05). Each of the above values is expressed as the mean ± S.D. RANKL: receptor activator of NFκB ligand; OPG: osteoprotegerin; M-CSF, macrophage colony-stimulating factor.