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Total proteins of DPSCs were extracted following the kit (KeyGEN, China) protocol. After protein denaturalization, the protein concentrations were measured by bicinchoninic acid (BCA) protein assays (Beyotime, China). Equal amount of each sample was segregated via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred to a nitrocellulose membrane. After blocking, the membranes were incubated with primary antibody: mouse anti-β-actin (ab8226, Abcam, 1: 1000) and mouse anti-OSX (sc-393325, Santa Cruz Biotechnology, 1: 1000). Then, the membranes were incubated with goat anti-mouse IgG-horseradish peroxidase (Santa Cruz Biotechnology) and detected with a chemiluminescent reagent kit (Millipore). The expression level of β-actin was normalized. A GS-700 imaging densitometer (Bio-Rad) was used for image analysis.
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Cells were lysed in radioimmunoprecipitation assaylysis buffer (Cell Signaling Technology, Inc. Danvers, MA, USA)supplemented with (https://ctbelize.com/activation-key/?patch=270) protease inhibitor (Roche Diagnostics, Basel,Switzerland). The concentration of total protein was determinedusing a bicinchoninic acid assay kit (Keygen Biotech).
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