Linkage analysis between Sp6 and AI in AMI rats. A. Genomic PCR products amplified by WT- (upper panel) or AMI- (lower panel) Sp6-specific primers. Sample numbers and the characteristics of the animals are denoted above the gel image. B. The color of the incisors was examined. Pups derived from the cross between F1 and AMI rats were sorted by the Sp6 genotype to determine the correlation between Sp6 and AI in AMI rats (Wt/Ami, n = 62; Ami/Ami, n = 56). C. Schematic diagram of focal gene localization in rat chromosome 10q31–10q32.1. D. Western blot analyses. COS7 cells were transfected with expression vector only (V) or with FLAG-tagged Wt- (W) or Ami- (A) cDNA. Samples were blotted with the indicated antibodies. E. Immunohistochemical analysis of incisors from newborn pups. Sections from secretory stage incisors of WT rats and those of the corresponding region from AMI rats were stained with the anti-Wt-SP6 antiserum (SP6) or preimmune rabbit serum (Pre). Signals were obtained from DAB (brown). Sections were counterstained with Hematoxylin. Scale bar: 100 mm. A, ameloblasts; O, odontoblasts; P, pulp.