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. 2017 Sep 6;1(10):e000406. doi: 10.3945/cdn.117.000406

FIGURE 3.

FIGURE 3

Osteoclast differentiation is suppressed by a polyphenolic fraction of dried plum in TNF-α–stimulated co-cultures. Primary calvarial osteoblasts were treated with 1,25-hydroxyvitamin D3 (10 nM) and dried plum polyphenolic fractions (10 μg/mL) and co-cultured with BMMCs. (A) Multinucleated TRAP-positive osteoclasts were quantified per well. (B) The relative mRNA abundance of Rankl, Opg, Nfatc1, and cFos was assessed with qRT-PCR by using Gapdh as a control. (C) In addition, multinucleated TRAP-positive osteoclasts were quantified after 10 d of co-culture in the presence of TNF-α (1 ng/mL). (D) After 6 d of treatment with dried plum polyphenolic fractions and TNF-α, the relative mRNA abundance of Rankl, Opg, Nfatc1, and cFos was assessed with Gapdh as a control. Values are means ± SEs. Experiments were repeated 2–3 times with 6 replicates/experiment, and data were analyzed by using ANOVA followed by Fisher's least significant difference test. Bars without a common letter are significantly different from each other, P < 0.05. BMMC, bone marrow mononuclear cell; CON, control; DP-Fr, dried plum fraction; Nfatc1, nuclear factor of activated T cells, cytoplasmic, calcineurin-dependent 1; Opg, osteoprotegerin; Rankl, receptor activator of NF-κB ligand; TRAP, tartrate-resistant acid phosphatase.