Fig. 3.

Association of rs1541160/rs522444 with expression of KIFAP3. (A and B) Total RNA was isolated from lymphoblastoid cell lines (A) or occipital cortex brain tissue harboring (B) either a CC or TT genotype for rs1541160. Relative expression of KIFAP3 was determined by real-time PCR. As shown, individuals harboring the CC genotype display decreased expression compared with individuals with the TT genotype. Error bars represent the 95% C.I. (C) The sequence of the KIFAP3 promoter region is shown. The arrow indicates the transcriptional start site. SNP rs522444 is indicated at the −25 position. The box represents the location of the putative Sp1 binding site. (D) The KIFAP3 promoter region and 5′ UTR (633 bp) were amplified from individuals harboring either the CC or TT genotype and subcloned upstream of the firefly luciferase gene. The schematic (not drawn to scale) represents the resultant constructs, which differ only at a single base pair located at rs522444. (E) The resultant constructs were transfected into SKN-AS cells and relative luciferase activity was measured. The error bars represent the 95% C.I. A promoterless vector yielded <1% relative activity. The construct containing the G allele displays higher luciferase activity relative to the C allele. *, P < 0.05; **, P < 0.01.