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Table 1.

Sequences of DNA probes and competitors used for the EMSA with HLH-2 and LAG-1

EMSA with HLH-2
 
 
DNA probe containing wild-type E-boxes
 
 
gaattcaaaaattccttaCACCTGtctttccacctttcccgctccccccgtccattaccactttgttaaccttgggggcacactagtaaccattacattaggatttacaaatcggaaCACCTGgtgcttggtgaattc
 
 
DNA probe containing mutated E-boxes
 
 
gaattcaaaaattccttaCTCCTAtctttccacctttcccgctccccccgtccattaccactttgttaaccttgggggcacactagtaaccattacattaggatttacaaatcggaaCTCCTAgtgcttggtgaattc
 
 
Wild-type competitor DNAs
 
Mutated competitor DNAs
 
1st: aaattccttaCACCTGtctttccacc 1st: aaattccttaCTCCTAtctttccacc 
2nd: caaatcggaaCACCTGgtgccttggt 2nd: caaatcggaaCTCCTAgtgccttggt 
EMSA with LAG-1
 
 
DNA probe containing wild-type LAG-1 binding sites
 
 
gaattctcgcgactCGTGGGAAaatgggcggaagggcacCGTGGGAAaatagttccaggaattc
 
 
Wild-type competitor DNAs
 
Mutated competitor DNAs
 
1st: taataatctctTCCCAaactacacctc 1st: taataatctctAACCAaactacacctc 
2nd: cctaactcctaTCCCAaattcgtacat 2nd: cctaactcctaAACCAaattcgtacat 
3rd: ttcttgtttctTCCCAcgtacagtttc 3rd: ttcttgtttctAACCAcgtacagtttc 
4th: ttacaaattctTCCCAgaacccaaaac 4th: ttacaaattctAACCAgaacccaaaac 
5th: gggatccgagaTGGGAaaagagactcg 5th: gggatccgagaTGGTTaaagagactcg 
6th: acagtcgctgaTGGGAaaatggataat 6th: acagtcgctgaTGGTTaaatggataat 
7th: tgctctcacagTGGGAaaattagattc 7th: tgctctcacagTGGTTaaattagattc 
8th: gcgaagatctgTGGGAaaatatacaga 8th: gcgaagatctgTGGTTaaatatacaga 
EMSA with HLH-2
 
 
DNA probe containing wild-type E-boxes
 
 
gaattcaaaaattccttaCACCTGtctttccacctttcccgctccccccgtccattaccactttgttaaccttgggggcacactagtaaccattacattaggatttacaaatcggaaCACCTGgtgcttggtgaattc
 
 
DNA probe containing mutated E-boxes
 
 
gaattcaaaaattccttaCTCCTAtctttccacctttcccgctccccccgtccattaccactttgttaaccttgggggcacactagtaaccattacattaggatttacaaatcggaaCTCCTAgtgcttggtgaattc
 
 
Wild-type competitor DNAs
 
Mutated competitor DNAs
 
1st: aaattccttaCACCTGtctttccacc 1st: aaattccttaCTCCTAtctttccacc 
2nd: caaatcggaaCACCTGgtgccttggt 2nd: caaatcggaaCTCCTAgtgccttggt 
EMSA with LAG-1
 
 
DNA probe containing wild-type LAG-1 binding sites
 
 
gaattctcgcgactCGTGGGAAaatgggcggaagggcacCGTGGGAAaatagttccaggaattc
 
 
Wild-type competitor DNAs
 
Mutated competitor DNAs
 
1st: taataatctctTCCCAaactacacctc 1st: taataatctctAACCAaactacacctc 
2nd: cctaactcctaTCCCAaattcgtacat 2nd: cctaactcctaAACCAaattcgtacat 
3rd: ttcttgtttctTCCCAcgtacagtttc 3rd: ttcttgtttctAACCAcgtacagtttc 
4th: ttacaaattctTCCCAgaacccaaaac 4th: ttacaaattctAACCAgaacccaaaac 
5th: gggatccgagaTGGGAaaagagactcg 5th: gggatccgagaTGGTTaaagagactcg 
6th: acagtcgctgaTGGGAaaatggataat 6th: acagtcgctgaTGGTTaaatggataat 
7th: tgctctcacagTGGGAaaattagattc 7th: tgctctcacagTGGTTaaattagattc 
8th: gcgaagatctgTGGGAaaatatacaga 8th: gcgaagatctgTGGTTaaatatacaga 

Sequences of DNA probes for HLH-2 and competitors for HLH-2 and LAG-1 (see Fig. 6) from the C. elegans egl-43 gene. The DNA probe sequence for LAG-1 EMSA has been used for EMSA with CBF-1 and Su(H) proteins, mammalian and D. melanogasterorthologs of LAG-1 (Zimber-Strobl et al.,1994). Sequences of the upper strand of DNA duplexes are shown. Bold characters represent wild-type or mutated E-boxes and LAG-1 binding sites.

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