Table 2. Sequences of double-stranded oligonucleotides used in EMSA analysis.
All oligonucleotides have a 3′-GGG overhang for labelling with [32P]dCTP with exceptions as follows: the AP-1 consensus and 2J2-A (−152/−103) oligonucleotides have a 5′-CTAG overhang on the sense strand and a 5′-GATC overhang on the antisense strand; the Stat5 antisense oligonucleotide has a 5′-G overhang. Mutated nucleotides are underlined. The AP-1 consensus sequence in the sense strand of the AP-1 consensus oligonucleotide, the Stat5 element in the Stat5 β-casein promoter oligonucleotides, and the c-Jun binding site at −105/−95 bp in the sense strand of probe 5 (−105/−86) are indicated in bold.
| Oligonucleotide | Sense/antisense | Sequence (5′–3′) |
|---|---|---|
| 2J2-A (−152/−103) | Sense | CTAGCCCGGGAATCCAGCGCCTGGCATCTTCGCAGGGTGCTGCGAAGGGGCGG |
| 2J2-A (−152/−103) | Antisense | GATCCCCGCCCCTTCGCAGCACCCTGCGAAGATGCCAGGCGCTGGATTCCCGGG |
| 2J2-B (−127/−79) | Sense | TCGCAGGGTGCTGCGAAGGGGCGGGCTGGGAGGCGGGGCACGGCTGGGAGGG |
| 2J2-B (−127/−79) | Antisense | TCCCAGCCGTGCCCCGCCTCCCAGCCCGCCCCTTCGCAGCACCCTGCGAGGG |
| 2J2-C (−102/−50) | Sense | CTGGGAGGCGGGGCACGGCTGGGAGCGAGGCGGGGCGGGGACCGTCGCCTGCTGGG |
| 2J2-C (−102/−50) | Antisense | AGCAGGCGACGGTCCCCGCCCCGCCTCGCTCCCAGCCGTGCCCCGCCTCCCAGGGG |
| 1 (−152/−128) | Sense | CCCGGGAATCCAGCGCCTGGCATCTGGG |
| 1 (−152/−128) | Antisense | AGATGCCAGGCGCTGGATTCCCGGGGGG |
| 2 (−137/−119) | Sense | CTGGCATCTTCGCAGGGTGGG |
| 2 (−137/−119) | Antisense | ACCCTGCGAAGATGCCAGGGG |
| 3 (−127/−106) | Sense | TCGCAGGGTGCTGCGAAGGGGCGGG |
| 3 (−127/−106) | Antisense | GCCCCTTCGCAGCACCCTGCGAGGG |
| 4 (−114/−97) | Sense | CGAAGGGGCGGGCTGGGAGGG |
| 4 (−114/−97) | Antisense | TCCCAGCCCGCCCCTTCGGGG |
| 5 (−105/−86) | Sense | GGGCTGGGAGGCGGGGCACGGGG |
| 5 (−105/−86) | Antisense | CGTGCCCCGCCTCCCAGCCCGGG |
| 6 (−94/−77) | Sense | CGGGGCACGGCTGGGAGCGGG |
| 6 (−94/−77) | Antisense | GCTCCCAGCCGTGCCCCGGGG |
| 7 (−85/−69) | Sense | GCTGGGAGCGAGGCGGGGGG |
| 7 (−85/−69) | Antisense | CCCGCCTCGCTCCCAGCGGG |
| 8 (−77/−60) | Sense | CGAGGCGGGGCGGGGACCGGG |
| 8 (−77/−60) | Antisense | GGTCCCCGCCCCGCCTCGGGG |
| 9 (−68/−48) | Sense | GCGGGGACCGTCGCCTGCTGGGGG |
| 9 (−68/−48) | Antisense | CCAGCAGGCGACGGTCCCCGCGGG |
| mt 5 (−105/−86; mt −105/−96) | Sense | AAAAAAAAAAGCGGGGCACGGGG |
| mt 5 (−105/−86; mt −105/−96) | Antisense | CGTGCCCCGCTTTTTTTTTTGGG |
| AP-1 consensus | Sense | CTAGTGATGAGTCAGCCGGATC |
| AP-1 consensus | Antisense | GATCGATCCGGCTGACTCATCA |
| Stat5 β-casein promoter | Sense | GGACTTCTTGGAATTAAGGGA |
| Stat5 β-casein promoter | Antisense | GTCCCTTAATTCCAAGAAGTCC |