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. 2021 Jan 5;22(1):449. doi: 10.3390/ijms22010449

Figure 4.

Figure 4

Inhibition of early osteoclastogenesis through TXA. (a) TRAP activity staining of WT bone marrow cells at day 7 of differentiation, cultured in the presence of M-CSF and RANKL and continuous exposure to vehicle or TXA at the indicated concentrations. Scale bars = 50 μm. The quantification of osteoclast numbers per viewing field is depicted on the left (Ocl.N./VF). (b) TRAP activity staining of WT bone marrow cells at day 7 of differentiation, cultured in the presence of M-CSF and RANKL, and exposure to vehicle or TXA at the indicated concentrations only at day 1 of osteoclast differentiation. The quantification of osteoclast numbers per viewing field is depicted on the left (Ocl.N./VF). (c) TRAP activity staining of WT osteoclasts at day 7 of differentiation, cultured in the presence of M-CSF and RANKL and exposed to vehicle or TXA at the indicated concentrations for 24 h. (d) MTT proliferation assay of WT osteoclasts at day 7 of differentiation stimulated with the indicated concentrations of TXA for 6 h. (e) qRT-PCR expression analysis for the indicated genes in bone marrow-derived osteoclasts at day 7, stimulated with TXA (1 mg/mL) during the entire course of cell differentiation. (f) qRT-PCR expression analysis for the indicated genes in bone marrow-derived osteoclasts at the indicated stages of cell differentiation, stimulated with TXA (1 mg/mL) for 6 h. (g) qRT-PCR expression analysis for the indicated genes in bone marrow-derived osteoclasts at day 5 of differentiation, stimulated with TXA (1 mg/mL) during the entire course of cell differentiation. For (a–g), n = 3–4 independent cultures per group were used. Data presented are means ± SD. Gene abbreviations: chloride channel 7 (Clcn7), nuclear factor kappa B subunit 1 (Nfkb1), receptor activator of nf-κb (Tnfrsf11a), tartrate-resistant acid phosphatase (Acp5), cathepsin K (Ctsk).