Oat Seedlings Extract Inhibits RANKL-Induced c-Fos/NFATc1 Transcription Factors in the Early Stage of Osteoclast Differentiation

Figure 2

Inhibitory effect of osteoclast differentiation in the early stages in the presence of oat seedlings’ extract (OSE). (a) Representation of the exposure schedule for OSE (100 μg/mL) treatment of the bone marrow-derived macrophages (BMMs) for various time periods (the indicated black arrow) in the presence of macrophage-colony stimulating factor (M-CSF; 30 ng/mL) and the receptor activator of nuclear factor κB ligand (RANKL; 10 ng/mL). (b) Formation of the tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells (MNCs) photographed under a light microscope (10x magnification). (c) Counting of the TRAP + MNCs (left panel) and the TRAP activity estimation (right panel). (d) Staining of the actin rings of mature osteoclast with the phalloidin-FITC. (e) Visualization of the TRAP + MNCs (top images) and resorption areas (bottom images) under a light microscope (10x magnification). The resorption areas were photographed by removing the cells. Results of one representative experiment out of three independent experiments yielding similar results are shown. (f) Measurement of the TRAP activity during osteoclast apoptosis on the slides (left panel) and quantification of the resorptive areas (%) by using the ImageJ program (right panel). N.S, not significant; (versus the control); , (versus the RANKL-treated control).

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