Table 1. QRT–PCR primers.

NCBI RefSeq	Gene	Forward primer (5′-3′)	Reverse primer (5′-3′)
NM_001080448	Eph-A6	TTTTTCTCCCAAGCCATTCA	ATGCCCAGTCCTTCCTTACC
NM_004428	ephrin-A1	GGTGACTGTCAGTGGCAAAA	GCACTGTGACCGATGCTATG
NM_001405	ephrin-A2	GCCTGCGACTGAAGGTGTA	CGGGCTGCTACACGAGTTAT
NM_004952	ephrin-A3	ACTCTCCCCCAGTTCACCAT	GTCCCGCTGATGCTCTTCT
NM_005227	ephrin-A4	ACTACATCTCGGTGCCCACT	CTGATGTGCCACTCTCTCCA
NM_001962	ephrin-A5	TTCATGATCGTGTTTTCGATG	GCTGGGTATCCTTGGTGTTT
NM_002046	GAPDH	TGCACCACCAACTGCTTAGC	GGCATGGACTGTGGTCATGAG

QRT–PCR primers were designed to verify gene expression of Eph-A6 and its ligands ephrin-A1 and -A4, as well as ephrin-A2, -A3, and -A5. These genes were amplified using a SYBR GreenER QRT–PCR kit (Invitrogen) and amplification was normalized relative to a GAPDH reference gene. QRT–PCR reactions were done in triplicate and repeated using cDNA from three different specimens.