Anti dspp

Tissue sections or cells were fixed, washed and blocked, and then incubated with the following primary antibodies: anti-Nestin (1/200; Abcam), anti-NF200 (1/200; Abcam), anti-MBP101 (1/200; Abcam), anti-Active β-catenin (1/200; Cell Signaling Technology), anti-DSPP (1/200; Zen Bioscience), anti-DMP1 (1/200; Santa Cruz Biotechnology), anti-CD31 (1/200; Zen Bioscience), and anti-VEGF (1/200; Abcam). The goat anti-rabbit 488 (1/200; Invitrogen) and goat anti mouse 488 (1/200; Invitrogen) secondary antibodies were used. Sections or cells were counterstained with DAPI, mounted with fluorescent mounting media and visualized using a fluorescence confocal microscope (Olympus FV1200, Olympus). Accordingly, quantitative analysis of the staining was conducted by measuring the average optical density using the ImageJ software. At least 3 replicates were performed.

For histology and immunohistochemistry analysis, sections of 6 µm were made for hematoxylin/eosin staining. Immunohistochemical staining was performed as previously described 35 (link). Primary anti-DSPP (1/200; Zen Bioscience), anti-DMP1 (1/200; Santa Cruz Biotechnology), and anti-OCN (1/200; Zen Bioscience) antibodies were used. Detection of crossreacted secondary antibodies was performed using the DAB kit (Gene Tech, China). Accordingly, quantitative analysis of the staining was conducted by measuring the average optical density using the ImageJ software. At least 3 serial sections from each sample were examined, and at least 3 samples were included in each group.