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. Author manuscript; available in PMC: 2011 Apr 1.
Published in final edited form as: Bone. 2010 Jan 4;46(4):1131–1137. doi: 10.1016/j.bone.2009.12.026

Figure 1.

Figure 1

a) Schematic of the FZD1 5′ UTR region. The relative locations of rs2232157 and rs 2232158 are shown. b) Quantitative RT-PCR analysis of FZD1. Relative expression level (ΔCt) was calculated using ΔCt of GAPDH as a reference. c). Western blot analysis of FZD1 protein in MG63 and SaOS-2 cells. Beta-actin was used as an internal control to demonstrate protein loading. d) Activity of the haplotype specific FZD1 promoters. The promoter activity for different FZD1 haplotypes were assessed by transfecting MG63 and SaOS-2 cell lines with luciferase reporter plasmids. Differences in activity were tested using the Kruskal-Wallis and Wilcoxon rank sum test. e) Allele specific binding to the rs2232157 specific probes by nuclear proteins. EMSA was carried out using radio-labeled probes for the two alleles of rs2232157 with nuclear extracts from both MG63 and SaOS-2 cells. The two major complexes are labeled with A and B. To ensure specificity of binding, the reaction was incubated with 50-fold excess unlabeled G and T probes (denoted by g* and t*, respectively). As a control, binding reaction with radio-labeled probe without nuclear extract was conducted (denoted as C).