Immunohistochemical detection of osteocalcin, DSP and MEPE after treatment with MEPE-ASARM peptides.

<p>SHED cell cultures were maintained in nonmineralizing (NM) or mineralizing (M) conditions in the absence or presence of 20 µM of either phosphorylated (p-ASARM) or nonphosphorylated (np-ASARM) ASARM peptide for 21 days. Immunohistochemistry for osteocalcin, DSP and MEPE was performed on methyl methacrylate sections of 21-day cultures. Osteocalcin immunostaining (top row) is strong in SHEDs (arrows) and nodules (arrowheads) in the M and M+p-ASARM conditions. Immunohistochemistry for DSP shows strong staining in mineralized nodules in both the M and M+np-ASARM conditions. MEPE immunostaining is found in SHEDs and nodules in mineralizing conditions the (M and M+np-ASARM conditions) but is particularly strong in the cultures treated with p-ASARM that do not mineralize (M+p-ASARM). The images presented are representative of all sections examined.