Method: The gene expression was analyzed using several mouse tissues by real time PCR. The expression and localization of Fam20a protein were examined using anti-Fam20a antibody by immunofluorescence-based staining and immunoprecipitation (IP) - Western Blot (WB) techniques in LS8 cells.
Result: Fam20a was highly expressed in lung, liver and tooth at mRNA levels. The expression was also confirmed in the LS8 ameloblast cell line. The IP-WB analysis was performed using cultured media and cell lysates from LS8 cells and the protein expression was clearly observed in the cell lysate fraction. Cell lysates from LS8 cells were further fractionated and the WB analysis demonstrated that the endogenous Fam20a protein was present in membrane fraction. The subcellular localization of endogenous Fam20a protein was detected at the cytoplasm.
Conclusion: We demonstrated the intracellular expression of FAM20A in LS8 ameloblasts. Our data suggest that FAM20A is a novel intracellular modulator of enamel formation/mineralization.
Supported by NIH grant DE019527 and Boston University School of Dental Medicine.
Keywords: Ameloblasts, Amelogenesis Imperfecta, Gene expression and Teeth
See more of: Craniofacial Biology