Table 2. PCR amplification, pyrosequencing, and RT-PCR primers used in measuring DNA methylation and mRNA levels of the S100A15 gene and its promoter regions.

Primers	Sequences
Region 1 for -583 CpG site
Forward PCR Primer	5′-GATTTATAGTATAGGGGATAAGAGTAAGGA
Biotinylated Reverse PCR Primer	5′-TCCCAAATTTACTCAACTAATAAACACAAC
Forward Sequencing Primer	5′-GGGGATAAGAGTAAGGAT
Region 2 for -423/−412 CpG site
Forward PCR Primer	5′-GGGTTGAGGAGGATGTAAGAGTAGT
Biotinylated Reverse PCR Primer	5′-CTCCCCCAATACTTCCACACATTT
Forward Sequencing Primer	5′-AAGAGTAGTAGGTGTTTT
Region 3 for -248 CpG site
Biotinylated Forward PCR Primer	5′-TGTTTATTAGTTGAGTAAATTTGGGAATAT
Reverse PCR Primer	5′-CAAACTCCCAAAAACAACATAATCTTATT
Reverse Sequencing Primer	5′-ACACATTTATTATAACAACCTATT
Region 4 for -56 CpG site
Forward PCR Primer	5′-ATGTTGTTTTTGGGAGTTTGAAAT
Biotinylated Reverse PCR Primer	5′-AACACACCCAAAAAACTATACTC
Forward Sequencing Primer	5′-ATTTTTTGATTTGTTATTATATATG
Region 5 for +21 CpG site
Forward PCR Primer	5′-AGGTTGAGTTTTATAAAGGATTGTTT
Biotinylated Reverse PCR Primer	5′-CCCAAACTTCTAAAACTTTATCCACAA
Forward Sequencing Primer	5′-ATAAAGGATTGTTTTTTGTTTAAAT
S100A15 LF RT-PCR
hS100A15LF-Forward	5′-ACGTCACTCCTGTCTCTCTTTGCT
hS100A15LF-Reverse	5′-TGATGAATCAACCCATTTCCTGGG
S100A15 SF real-time RT-PCR
hS100A15SF-Forward	5′-CAAGTTCCTTCTGCTCCATCTTAG
hS100A15SF-Reverse	5′-AGCCTTCAGGAAATAAAGACAATC
GAPDH real-time RT-PCR
Forward	5′-GAAGAGCCAAGGACAGGTAC
Reverse	5′-CAACTTCATCCACGTTCACC