Table 1.

Primer sequences and PCR conditions

Gene	PCR	Forward primer sequence (5′–3′)	Reverse primer sequence (5′–3′)	Annealing temperature	PCR cycles
NELL1	Quantitative RT-PCR	ATGGCCAGGTGTGGACCTTG	TCTGGTGTCACATTCTGGGC	60	40
NELL2	Quantitative RT-PCR	CCACTTAAGTCGGCTCTTGC	ATGGTCTGGTCCTTGCACTC	60	40
GAPDH	Quantitative RT-PCR	GTCTCCTCTGACTTCAACAGCG	ACCACCCTGTTGCTGTAGCC	60	40
NELL1	MSP-Methyl.	GATTTCGATTTCGGTGTTTC	CTACGTTCGTCTAAAAACCCG	60	35
NELL2	MSP-Methyl.	CGTTTTATTTGGTAGGGGGC	TTCCCTACCGAAAACGAAA	60	35
NELL1	MSP-Unmethyl.	TGGGATTTTGATTTTGGTGTTTT	ACTACATTCATCTAAAAACCCAAAC	55	40
NELL2	MSP-Unmethyl.	TTATGTTTTATTTGGTAGGGGGT	AATTCCCTACCAAAAACAAAAC	63	35
NELL1	Bisulfite sequencing	TTATAGGGGTTTAATAGGAGAGAGG	CCTAACTTAAAACCR*CCAAACTTACTAA	55	35
NELL2	Bisulfite sequencing	TGTTTTTTGTTGTATGTTGGTTT	AAAAAAAAAAACCTCCCCAAAC	55	35

^*R(A+G). Methyl., methylated; MSP, methylation-specific PCR; Unmethyl., unmethylated.