TABLE 1.
Name | Direction | Sequence |
---|---|---|
EMSA probes | ||
GS-ECR | Forward | 5′-gatcCTTAGTTATTCATTTCTGCAGAACTAATTTTTAACCTAG-3′ |
GS-ECR | Reverse | 5′gatcCTAGGTTAAAAATTAGTTCTGCAGAAATGAATAACTAAG-3′ |
GS-ECR_mut | Forward | 5′-gatcCTTA-GGGGTTCAGGTCTGCAGAACGGATTTTGGGCCTAG-3′ |
GS-ECR_mut | Reverse | 5′-gatcCTAGGCCCAAAATCCGTTCTGCAGACCTGAACCCCTAAG-3′ |
GS-HNF1_cons | Forward | 5′-gatcCCAGGTTAATGATTAACCCA-3′ |
GS-HNF1_cons | Reverse | 5′-gatcTGGGTTAATCATTAACCTGG-3′ |
ChIP primers | ||
ChIP-ECR | Forward | 5′-CTGGCTCGTGGGTAAGAATTGTCTC-3′ |
ChIP-ECR | Reverse | 5′ATGTCCAAGGCAATGCTAGGTTAAA-3′ |
ChIP-Intron2 | Forward | 5′-GGCAGCGAGATCGAAGGACAACTGT-3′ |
ChIP-Intron2 | Reverse | 5′-TCTCCTGCCTTCGGGTTTTCTCCAA-3′ |
Primers used in the generation of the reporter gene and the HNF1-overexpressing constructs | ||
OCT1 Promoter | Forward | 5′-TGCACAGAGAGAGAAACCAAAAGTC-3′ |
OCT1 Promoter | Reverse | 5′-GCCAGCTCGAGATGTCTCCCTCAGAGATCTTTG-3′ |
ECR | Forward | 5′-TCCTTGGATCCCCAGCTCCTCCTCCAAACT-3′ |
ECR | Reverse | 5′-CCCGAGTCGACCTCATGATCTTAGCACCTAGCCTT-3′ |
HNF1A | Forward | 5′-CCTGTGGATCCGAGCCATGGTTTCTAAACTGA-3′ |
HNF1A | Reverse | 5′-AGCTTATCTAGAGTGGTTACTGGGAGGAAGAGG-3′ |
HNF1B | Forward | 5′- CTTTTTCCGGATCCTTGGAAAATGGTGTCCAA-3′ |
HNF1B | Reverse | 5′- GTGGTGTCTAGAGGCATCACCAGGCTTGTAGA-3′ |
Primers used for site-directed mutagenesis of the ECR1 Binding Sites | ||
HNF1A(A)_f | Forward | 5′-TTGGGGAATCAATCTTAGGGGTTCAGGTCTGCAGAACTAATTTTT A-3′ |
HNF1A(A)_r | Reverse | 5′-TAAAAATTAGTTCTGCAGACCTGAACCCCTAAGATTGATTCCCCAA-3′ |
HNF1A(B)_f | Forward | 5′-GGTTCAGGTCTGCAGAACGGATTTTGGGCCTAGCATTGCCTTGGAC-3′ |
HNF1A(B)_r | Reverse | 5′-GTCCAAGGCAATGCTAGGCCCAAAATCCGTTCTGCAGACCTGAACC-3′ |
ChIP, chromatin immunoprecipitation. The unspecific sequences used in the radioactive labeling of the EMSA probes are given in lowercase letters. Mutated nucleotides are shown in boldface, and the artificially introduced restriction sites are underlined. The ATG start codons of HNF1α and HNF1β are given in italics.