Table 1.
A.PIII deletion primers | Primer sequence (5’ – 3’) |
---|---|
PIII-XhoI forward | CGGCTCGAGACTGTTTTGCCTCCAAGCAAGGA |
PIII-HindIII reverse | ACCAAGCTTTTTTCAGTGACAGGTAAAGCTGTTTCC |
-386-PIII forward | CGGCTCGAGTAGTGAAGAAAGAGTGAACAAG |
-398-PIII forward | CGGCTCGAGGACTCCTCCTCTAGTGAAGAA |
B. PIII site-directed mutagenesis primers | Primer sequence (5’ – 3’)* |
AP1 forward reverse | CGCCTTCCTGCTCTGTCTAACTAACTCCTCTCCTGCGTTCTGG CCAGAACGCAGGAGAGGAGTTAGTTAGACAGAGCAGGAAGGCG |
Sp1A forward reverse | GCATGTTGCAACACTGACTACTCATCTAGTGAAGAAAGAG CTCTTTCTTCACTAGATGAGTAGTCAGTGTTGCAACATGC |
Sp1B forward reverse | GAGCTGGGCTTTCCACGCATTCCTGCTCTGTCTCACTC GTGAGACAGAGCAGGAATGCGTGGAAAGCCCAGCTC |
C/EBP forward reverse | CCAGCAAGGAACATGCATGCTACAACACTGACTCCTCCTCTAG CTAGAGGAGGAGTCAGTGTTGTAGCATGCATGTTCCTTGCTGG |
Double forward reverse | CAGCAAGGAACATGCATGTTACAATACTGACTACTCATCTAG CTAGATGAGTAGTCAGTATTGTAACATGCATGTTCCTTGCTC |
Triple forward reverse | CAGCAAGGAACATGCATGTTACAATACTGACTACTCATCTAG CTAGATGAGTAGTCAGTATTGTAACATGCATGTTCCTTGCTC |
* Template for all site-directed mutagenesis constructs is -439-PIII deletion mutant; template for triple had Sp1B site already mutated. | |
C. Gel shift oligos | Primer sequence (5’ – 3’) |
Sp1A forward reverse | ATGTTGCAACACTGACTCCTCCTCTAGTGAAGA TCTTCACTAGAGGAGGAGTCAGTGTTGCAACAT |
Sp1A mutated forward reverse | ATGTTGCAACACTGACTACTCATCTAGTGAAGA TCTTCACTAGATGAGTAGTCAGTGTTGCAACA |
Sp1B forward reverse | GAGCTGGGCTTTCCCCGCCTTCCTGCTCTGTCT AGACAGAGCAGGAAGGCGGGGAAAGCCCAGCTC |
Sp1B mutated forward reverse | GAGCTGGGCTTTCCACGCATTCCTGCTCTGTCT AGACAGAGCAGGAATGCGTGGAAAGCCCAGCTC |
C/EBP forward reverse | AGGAACATGCATGTTGCAACACTGACTCCT AGGAGTCAGTGTTGCAACATGCATGTTCCTTGC |
C/EBP mutated forward reverse | GCAAGGAACATGCATGTTACAATACTGACTCCT AGGAGTCAGTATTGTAACATGCATGTTCCTTGC |