Protein Expression Analysis in Osteogenesis

Whole cell lysates and western blot analyses were performed as described previously44 (link). The primary antibodies used were as follows: Runx2 rabbit monoclonal antibody (1:2000) (Epitomics, Burlingame CA), anti-Sp7/Osterix rabbit polyclonal antibody (1:5000) (Abcam, UK), anti-KAT3B/p300 antibody (1:2000) (Abcam), and anti-acetyl lysine antibody (1:1000) (Abcam). Membranes were incubated for 1 h at room temperature with the primary antibody in 5% milk followed by another incubation with a horseradish peroxidase-conjugated secondary antibody. The signals were detected using the Super Signal West substrate (Thermo Fisher Scientific, USA). Densitometry analyses of the western bands were performed using the Tanon Imaging software67 (link).