Table I.

Summary of the primer sequences, PCR product size obtained and annealing temperature used for MSP assays.

Gene	Primer	Sequence (5′-3′)	Annealing temperature (°C)	PCR product size (bp)
SFRP2	M	F: 5′-TTTTTGTAGGGGCGTTTTTATAAC-3′	54	163
		R: 5′-TATCGATATACTCCCCAATACCG-3′
	U	F: 5′-AGATTTTTGTAGGGGTGTTTTTATAAT-3′	52	163
		R: 5′-ACCTATCAATATACTCCCCAATACCA-3′
GATA4	M	F: 5′-GTCGGGATAGTTTTTCGTTC-3′	52	134
		R: 5′-CGATTTAAAACCGACAATCA-3′
	U	F: 5′-AAGGTTGGGATAGTTTTTTGTTT-3′	50	134
		R: 5′-TCCCAATTTAAAACCAACAATCA-3′
GATA5	M	F: 5′-TTAGAAATCGAGGAAATCGC-3′	54	133
		R: 5′-GTAAACCCCCTCGTTACGTA-3′
	U	F: 5′-TGTTTAGAAATTGAGGAAATTGT-3′	48	133
		R: 5′-CCCATAAACCCCCTCATTACATA-3′
NDRG4	M	F: 5′-GGAGTTTAAATAAAGATTACGGTAGC-3′	50	103
		R: 5′-ATACGCTACGAAACCCTACC-3′
	U	F: 5′-GGGAGTTTAAATAAAGATTATGGTAGT-3′	48	103
		R: 5′-AATACACTACAAAACCCTACC-3′
VIM	M	F: 5′-AGGAAAGTATAAATTTCGGGTGC-3′	52	173
		R: 5′-ATAAACGACGTCTTTCACCCTTAC-3′
	U	F: 5′-AAAAGGAAAGTATAAATTTTGGGTGT-3′	48	173
		R: 5′-TATAAACAACATCTTTCACCCTTACCT-3′

PCR, polymerase chain reaction; MSP, methylation-specific; bp, base pair; SFRP2, secreted frizzled-related protein 2; GATA4/5, GATA binding protein 4/5; NDRG4, N-Myc downstream-regulated gene 4; VIM, vimentin; M, methylated; U, unmethylated; F, forward; R, reverse.