The constructs of 5′proximal promoter regions of the mouse TH gene with or without potential binding sites of FoxO1 (TH_long, from −3,002 to −1, TH_medium, from −696 to −1 and TH_short, from −142 to −1) were amplified by PCR and cloned upstream of a luciferase reporter gene in pGL3-basic vector via KpnI and NheI sites for TH_long and TH_short constructs and via XhoI and KpnI sites for TH_medium construct. The PCR primers used include forward, 5′-GGGGGTACCGCTTCCCAGCTACTCC-3′ and reverse, 5′-CCCGCTAGCAAGCTGGTGGTCCC-3′ for TH_long construct; forward, 5′-GGGGGTACCGGGAGATGCCAAAGGC-3′ and reverse, 5′-CCCCTCGAGAAGCTGGTGGTCCCGA-3′ for TH_medium construct; forward, 5′-GGGGGTACCCCTCAGGCACAGCA-3′ and reverse, 5′-CCCGCTAGCAAGCTGGTGGTCCC-3′ for TH_short construct.
