RNA sequencing alignments for I.1, II.1, and II.2 at the variation position c.2500 cytosine (C) > thymine (T) (red box) and arginine (Arg) 834* (dashed box) are shown (A). Reads per reference (Ref) C allele and alternate (Alt) T allele are shown to the right. RFLP mapping schematic showing fragments generated from undigested or DdeI-digested amplicons from esophageal DHTKD1 transcripts generated from the C vs. T allele (B, upper panel). Gel electrophoresis after RFLP mapping performed on individuals I.1, II.1, and II.2 and normal control individual (NL) homozygous for the C allele (B, lower panel).+ CTL, positive control.