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Figure 1. Characterization of ameloblast-lineage cells by immunohistochemistry and real-time PCR. (A) DSP immunostaining showed strong staining by most ameloblast-lineage cells. Select cells stained positive for KLK-4 (B) and TNF-
(C). Controls showed only nuclear staining (D). Scale bar: 20 µm. (E) Quantitative PCR showed mRNA expression of TNF, DSPP, and KLK4 in pooled ameloblast-lineage cells exposed to either 0 or 10 µM F (RQ = relative quantity). Average (SD) values of RQ of the gene tested: (KLK4) RQctrl = 1.21 ± 0.37, RQF = 0.90 ± 0.31; (TNF) RQctrl = 0.81 ± 0.51, RQF = 1.06 ± 0.71; (DSPP) RQctrl = 0.87 ± 0.63, RQF = 0.43 ± 0.14 (n = 3). (F) DSPP was up-regulated in the youngest (group 3) cells derived from second molars exposed to F. Cells from incisors (group 1) or canines and first molars (group 2) were not affected. (G) TNF was up-regulated in the youngest F-exposed cells.
