Df6824

Sections were dewaxed in xylene and then rehydrated through an ethanol concentration gradient. Tissue samples were then deactivated with 3% H 2 O 2 . Antigen retrieval was performed using trypsin (Cat. #T8150, Solarbio, Beijing, China), followed by blocking the sections with 5% goat serum (Cat. #AR0009, BOSTER, Pleasanton, CA, USA) for 1 hour. Subsequently, sections were incubated overnight at 4 °C with specific antibodies against βcatenin (1∶500, Cat. #PK02151, Abmart, Shanghai, China), RUNX2 (1∶200, Cat. #sc-390 351, Santa Cruz Biotechnology, Dallas, TX, USA), OSX (1∶500, Cat. #ab209484, Abcam, Cambridge, UK), OPG (1∶500, Cat. #DF6824, Affinity, Cincinnati, OH, USA), NFATc1 (1∶200, Cat. #sc-7294, Santa Cruz Biotechnology, Dallas, TX, USA), c-Fos (1∶500, Cat. #66590-1-Ig, Proteintech, Rosemont, IL, USA), CTSK (1∶200, Cat. #sc-48353, Santa Cruz Biotechnology, Dallas, TX, USA), and RANKL (1∶500, Cat. #bs-20647R, Bioss, Woburn, MA, USA). Afterward, the sections were incubated with HRP-conjugated secondary antibodies at room temperature for 1 hour, followed by counterstaining with hematoxylin and sealing with neutral resin. Immunoreactivity was detected using an optical microscope (Olympus, Tokyo, Japan).