Figure 2. Results of sequencing analysis, PCR analysis, and single-base primer extension assay. A: Nucleotide and amino acid sequence of TACSTD2. Arrowheads indicate the site of the c.675C>A and c.840_841insTCATCATCGCCGGCCTCATC nucleotide changes. Note that the amino acids in bold italic type are of the transmembrane domain. B: Computationally-predicted domain structure of the TACSTD2 protein with mutations of previous reports and this report (*). SS: signal sequence; EGF: EGF-like repeat; TY: thyroglobulin type I repeat; TM: transmembrane domain; PIP2: PIP2 binding sequence. C: Results of sequencing analysis of TACSTD2 in a normal volunteer (upper) and in proband A or B (lower). The underlined nucleotides indicate the inserted 20-base sequence between the 840th and 841st nucleotide positions of TACSTD2. Note that the presented sequence is in a reverse direction. D: Results of sequencing analysis of TACSTD2 in a normal volunteer (upper) and in proband C (lower). Arrowheads indicate the site of the c.675C>A mutation. Note that the presented sequence is in a reverse direction. E: Results of PCR analysis to examine the difference in length between the normal and insertion-bearing alleles in the families of the proband A (left) and proband B (right). The upper bands indicate the PCR product derived from the insertion-bearing alleles while the lower bands indicate the PCR product from the normal alleles. Note that the sister of proband A was not examined. F: Results of 1-base primer extension analysis for the 675th nucleotide of TACSTD2 in the normal volunteer (left) and the proband C (right). Black indicates C and green indicates A. Note that the presented data was produced by the forward primer.