Tnf α sc 52746

Lipopolysaccharide (LPS) treatment was selected as a stimulus to provide the theoretical basis for the establishment of an LPS-induced inflammation model. LPS powder (Sigma-Aldrich), St. Louis, MO, USA) was dissolved in Hank’s Balanced Salt solution without Ca²⁺ and Mg²⁺ (Thermo Scientific, Rockford, IL, USA) to obtain a stock solution of 1 mg/mL LPS and stored at −20 °C for future use. Cells were stimulated with 1 µg/mL LPS for 24 h and the protein expression of TNF-α (sc-52746) (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) was assessed by Western blot to confirm the inflammatory induction of LPS treatment on hBMMSCs.

ASTX (CAS No. 472-61-7) and STZ (CAS No. 18883-66-4) were purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA), while fetal bovine serum (FBS) was obtained from HyClone Laboratories, Inc. (Logan, UT, USA). Antibodies specific to Nrf2 (BS1258) and receptor activator of nuclear factor (NF)-κB ligand (RANKL; ALX-804-243) were purchased from Bioworld Technology, Inc. (St. Louis Park, MN, USA) and Enzo Life Sciences, Inc. (Farmingdale, NY, USA), respectively. Antibodies specific to interleukin-1β (IL-1β; sc-52012), interferon-γ (IFN-γ; sc-57208), tumor necrosis factor α (TNF-α; sc-52746), and β-actin (sc-47778) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 2′,7′-Dichlorodihydrofluorescein-diacetate (DCF-DA), and anti-osterix (ab209484), anti-osteopontin (OPN; ab8448), anti-heme oxygenase 1 (HO-1; ab13248), anti-bone morphogenetic protein 2 (BMP2; ab214821), and anti-γ-H2AX (ab26350) antibodies were purchased from Abcam (Cambridge, UK). Unless otherwise specified, other reagents were purchased from Sigma-Aldrich Co. LLC, while laboratory consumables were from Falcon Labware (Becton-Dickinson, Franklin Lakes, NJ, USA).