Quantitative PCR Protocol for Gene Expression

Quantitative PCR was performed using the Superscript III One Step kit (Invitrogen Corp) in a Roche Lightcycler 480 (Roche Diagnostics). Standard curves were generated for each primer set, and samples fitted to the linear portion of the curve. Additionally, PCR products were run on DNA gels to verify single products. Primer sequences used were described previously and were as follows: FXNFWD 5′- ATCTTCTCCATCCAGTGGACCT-3′ and FXNREV 5′- GCTGGGCATCAAGCATCTTTT-3′; ARE16223FWD 5′- CCTGCCGTACTCAGTCCTTC-3′ and ARE16223REV 5′- CCACTCGGCTGTACTGTCTG-3′; NQO1FWD 5′-CCCTTTTAGCCTTGGCACGAAA-3′ and NQO1REV 5′- TGCACCCAGGGAAGTGTGTTGTAT-3′; HO1FWD 5′ CCCTGCTGAGTAATCCTTTCCCGA-3′ and HO1REV 5′- ATGTCCCGACTCCAGACTCCA-3′ (67 (link)).