Pgl2 basic luciferase reporter vector

Different region of Rad51 promoter sequences (transcription start site as +1; −700 to +300; −366 to +300; −133 to +300; −73 to +300; −13 to +300; +1 to +300; +43 to +300) was cloned into pGL2-basic luciferase reporter vector (Promega, Madison, WI, USA).

To construct the FAS-luc reporter plasmid, a 718 bp fragment (−668 to +51) of the human FAS promoter region was amplified by PCR using human genomic DNA as template (See Supplementary Table S1 for primer sequence information). The PCR product was digested with KpnI and HindIII, and inserted into the pGL2-basic luciferase reporter vector (Promega, Madison, WI, USA).

The fragments containing the potential HRE sites (5′-G/ACGTG-3′) identified from the LIF promoter (A: 582bp~-578bp; B: 1123bp~-1119bp; C: 1769bp~-1765bp; D: 2791bp~2787bp) were amplified by following PCR primers. For HRE-A, forward primer: 5′ CGG GGT ACC AGC ACC CGG AGG AGG AAA CTG G 3′, reverse primer: 5′ CCG CTC GAG CGG GGC CGG CGT GGA CTT 3′; for HRE-B, forward primer: 5′ CGG GGT ACC GGC TGG GCA GGT CTG GGA TGT GG 3′, reverse primer: 5′ CCG CTC GAG GTG GGG AGC CGG GCA AAA TGA GC 3′; for HRE-C, forward primer: 5′ CGG GGT ACC CGG GCG ACG GGG GTT TG 3′, reverse primer: 5′ CCG CTC GAG TTA CTG CTC CTA TAC ACT TGC TCT GGG G 3′; for HRE-D, forward primer: 5′ CGG GGT ACC GCC ATG CCC TTC GCC CTC TCA 3′, reverse primer: 5′ CCG CTC GAG TGC CAT CCA ACC CAT CAC TGC TCT 3′. A HRE site in the VEGF promoter (from −1080bp to −874bp upstream of transcription initiation site) was amplified using following primer set: forward: 5′-CGG GGT ACC CCT CAG TTC CCT GGC AAC ATC TG-3′, reverse: 5′-CCG CTC GAG GAA GAA TTT GGC ACC AAG TTT GT-3′. Each of forward primers and reverse primers contains a KpnI site and an XhoI site, respectively. These PCR fragments were cloned into pGL2-Basic Luciferase Reporter Vector (Promega) at KpnI-XhoI restriction sites. All constructs were confirmed by DNA sequencing.