β tublin

Automated immunoblot was carried out with Simple Wes (Protein Simple, USA) as described by the manufacturer. In brief, 1.5 μg total protein (cell lysate or exosome specimens) was loaded into a Wes assay plate, alongside antibody diluent (Protein Simple); primary antibodies targeting DSP (Santa Cruz, USA), DMP1, CD9, CD63, p-NFκΒ, IκB-a, Myd88, TLR4, IKBKB, BMP2, BMPR1, TGFβ1, TGFβ3, TGFR1, p-Smad2/3, p-Smad1/5/9, Smad4, β-actin, and β-Tublin (Affinity, USA); and anti-rabbit/mouse secondary antibodies (Protein Simple) and streptavidin-HRP. The compass software (Protein Simple) was employed for data analysis.

Total protein from DPSCs was obtained with RIPA (Cell Signaling, USA). Automated immunoblot was carried out with Simple Wes (Protein Simple, USA) as directed by the manufacturer. In brief, 2.5 μg total protein samples were mixed with the standard fluorescent master mix and added to prefilled Wes assay plates, alongside antibody diluent (Protein Simple); primary antibodies targeting BMP2 (Affinity, USA), BMPR1 (Affinity), p-Smad1/5/9 (Affinity), RUNX2 (Affinity), DSP (Santa Cruz, USA), DMP-1 (Genetex, USA), ERK, p-ERK, JNK, p-JNK, p38, p-p38 (Cell Signaling), and β-Tublin (Affinity, USA) were probed; anti-rabbit and anti-mouse secondary antibodies conjugated to Streptavidin-HRP (Protein Simple) were used for detection. The compass software (Protein Simple) was utilized for analysis.