Table 1. Summary of EMSA results from all 16 SNPs tested.
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Protein binding | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + | + |
Allele differences | - | - | + | + | + | - | + | + | + | + | + | + | + | + | + | + |
Competition assay | n/a | n/a | + | + | + | n/a | + | + | + | + | + | + | + | + | + | + |
Both alleles of all 16 SNPs showed a shift, indicative of protein binding with both HEK293 and SK-N-SH nuclear extracts (top row, see S4 Fig). SNPs are in columns 2 through 17 labeled as follows: 1.rs34382810, 2.rs7965923, 3.rs769087, 4.rs1006737, 5.rs2159100, 6.rs12315711, 7.rs11062170, 8.rs4765905, 9.rs758170, 10.rs2370414, 11.rs10774035, 12.rs10774036, 13.rs10744560, 14.rs12311439, 15.rs1024582, 16.rs4298967. SNPs rs34382810, rs7965923, and rs12315711 (1,2 and 6) did not show allele-specific binding with either type of nuclear extract (middle row), and were therefore not tested with a competition EMSA (bottom row). Competition EMSAs with SK-N-SH nuclear extracts for both alleles of all SNPs, which showed consistent differences in both cell lines, confirmed the allele-specificity observed in the standard EMSAs. “n/a” indicates not assayed.