Figure 2

Effect of the 5' UTR haplotype on reporter protein expression. A. Schematic representation of reporter gene constructs; details of construction provided in Methods and Additional file 1. Arrows indicate SNP sites in hBD1 5' UTR. Bent arrows indicate direction of translation. Light grey bars represent 5' UTR. Construct names are given at left, the SNP haplotypes comprise the latter portion of the name. B. and C. CAT expression in COS-7 cells transiently transfected with constructs containing hBD1 (or CMV) 5' UTR. CAT expression is normalized to β-galactosidase. (B) CAT activity measured by radioactive enzyme activity assay. Mock transfected (no DNA added) CAT values were subtracted and CAT activity was normalized to β-galactosidase activity obtained by ELISA enzyme activity assay. (C) The quantity of CAT protein was measured by an antibody-based ELISA and normalized to β-galactosidase protein. Black bars in B and C represent constructs containing hBD1 5' UTR with the -44 G allele. Values given are averages and standard errors from 3 or more experiments. Statistically significant differences are indicated with asterisks compared to pGEM-GGG. *, p < 0.05; ** p < 0.01; *** p < 0.001.