Table 1. Primer Sequences, PCR Product Sizes, and Annealing Temperatures Used in MSP Assays.

	PCR product size (bp)	Annealing temperature (℃)
SFRP2
M S: GGGTCGGAGTTTTTCGGAGTTGCGC	138	62
A: CCGCTCTCTTCGCTAAATACGACTCG
U S: TTTTGGGTTGGAGTTTTTTGGAGTTGTGT	145	50
A: AACCCACTCTCTTCACTAAATACAACTCA
BMP3
M S: GTTTGGAGTTTAATTTTCGGTTTC	179	54
A: ATAACTTCGATCTCTCTCCCTACG
U S: GGTTTGGAGTTTAATTTTTGGTTTT	178	54
A: AACTTCAATCTCTCTCCCTACACC
NDRG4
M S: TTTAGGTTCGGTATCGTTTCGC	110	61
A: CGAACTAAAAACGATACGCCG
U S: GATTAGTTTTAGGTTTGGTATTGTTTTGT	105	61
A: AAAACCAAACTAAAAACAATACACCA
TFPI2
M S: ATTTTTTAGGTTTCGTTTCGGC	118	57
A: GCCTAACGAAAAAAAATACGCG
U S: TTAGTTATTTTTTAGGTTTTGTTTTGGT	105	57
A: AAAAACACCTAACAAAAAAAAATACACA

MSP, methylation-specific PCR; SFRP2, secreted frizzled-related protein 2; M, methylated; S, sense; A, antisense; U, unmethylated; BMP3, bone morphogenetic protein 3; NDRG4, N-myc downstream-regulated gene 4; TFPI2, tissue factor pathway inhibitor 2.