Vericiguat Modulates Osteoclast Differentiation and Bone Resorption via a Balance between VASP and NF-κB Pathways

Figure 4

Vericiguat dually regulated RANKL-triggered osteoclast-specific gene expression in osteoclast formation. (a) The relative mRNA expression of osteoclast marker genes (c-Fos, NFATc1, TRAP, CTSK, CTR, and DC-STAMP) in BMMs treated with RANKL+MCSF, as well as indicated concentrations of Vericiguat, for three days was quantified by qRT-PCR (). Values are presented as the . indicated a comparison with the control group (Vericiguat, 0 nM): , , , and . (b, c) Western blot analysis results on the protein expression levels of OC-related markers including c-Fos and NFATc1 in BMM cells stimulated with RANKL for 5 d without or with Vericiguat with indicated concentrations and quantitative results (). indicated a comparison with the control group (Vericiguat, 0 nM): , , , and . (d, e) The cytoplasmic and nuclear fractions of the BMMs treated with 30 ng/mL M-CSF, 50 ng/mL RANKL, and Vericiguat (500 nM and 8 μM) for 0, 2, and 4 d, respectively, were analyzed by western blotting and quantitative results. GAPDH and histone H3 were used as nuclear and cytoplasmic loading controls, respectively (). indicated a comparison with the control group (Vericiguat, 0 nM) regarding the protein expression of NFATc1 within the cytosol at day 4 after stimulation: and . # indicated a comparison with the control group (Vericiguat, 0 nM) regarding the protein expression of NFATc1 within the nucleus at day 4 after stimulation: ^####. (f) Immunofluorescence results of the nuclear translocation of NFATc1 at day 4 after stimulation showed that Vericiguat at low concentration promoted the nuclear translocation of NFATc1, whereas high-concentration Vericiguat suppressed this effect. ns: not statistically significant. .

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