To test whether the major and minor alleles of the upstream region of CHRNB3 might affect gene expression, the CHRNB3 reporter constructs were evaluated in three cell types: HEK293T, P19, and SH-SY5Y. In the HEK293T cells, luciferase expression levels from the CHRNB3 constructs were not significantly higher than control plasmids containing no insert, suggesting these cell types do not contain the transcriptional machinery to express CHRNB3. In the P19 embryonal carcinoma cells, a one-way ANOVA including all five constructs yielded evidence for a significant difference between the constructs (F=2.87, p=0.018). However, none of the pair-wise tests for differences using Tukey’s post hoc test were significant (all p>0.08). The patterns of expression are shown in Figure 3a, which suggest a step-wise difference in expression between the three long constructs. A post hoc ANOVA of only these three constructs and the control revealed evidence for differences between the common and minor constructs. The overall ANOVA F statistic was 3.95 (p=0.012), with significant differences between the control and common haplotypes (p=0.048) and between the common and minor haplotypes (p=0.023). Figure 3a shows the
