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2 protocols using recombinant mouse s100a4

1

Osteoclastogenesis Signaling Pathway

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Recombinant M-CSF and RANKL were purchased from PeproTech. Recombinant mouse S100A4 was purchased from Prospec. Recombinant OPG was purchased from R&D Systems. Antibodies against vimentin and c-Fos were purchased from Santa Cruz. Antibodies against N-cadherin and snail2 were obtained from Cell Signaling Technology. Antibodies for RAGE (DD/A11) were from Millipore. Anti-S100A4 antibody was purchased from Abcam. Anti-β-catenin antibody was purchased from Invitrogen. Antibodies for NFATc1 (7A6), E-cadherin, and hHLA were purchased from BD Pharmingen. Anti-β-actin antibody (AC-74) and the leukocyte acid phosphatase kit (for TRAP staining) were purchased from Sigma-Aldrich. Lipofectamine for siRNA transfection was purchased from Life Technologies. siRNA oligonucleotides and shRNA lentiviral particles were purchased from Santa Cruz. Dentin slices were purchased from Immunodiagnostic Systems.
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2

Regulation of Osteoblast Differentiation

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Recombinant mouse S100A4 was purchased from Prospec (East Brunswick, NJ, USA). Phosphospecific antibodies against IκB kinase (IKK)α/β (Ser176/180), IκB (Ser32), p65 (Ser536), IKKα/β, IκB, and p65 were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibodies against osterix, laminB, and p65 were acquired from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-β-actin (AC-74) was purchased from Sigma-Aldrich (St. Louis, MO, USA), anti-Runx2 from MBL International (Woburn, MA, USA), the alkaline phosphatase (ALP) assay kit from Takara Bio Inc. (Fukui, Japan). β-Glycerophosphate, cetylpyridium chloride, and ascorbic acid from Sigma-Aldrich, Bay11-7082 from Alexis Biochemicals (Grunberg, Germany), and the Cell Counting Kit-8 (CCK-8) from Dojindo (Kumamoto, Japan).
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