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Figure S1. (A) Electrophoretic mobility shift assay (EMSA) showing absence of specific binding to SNP1 barring (B) or wild-type (N) ds-oligonucleotide. The last four lanes show binding to a probe harbouring a perfect E2F-1 binding site (PE). (B) EMSA showing absence of significant differences in binding to SNP2 barring (B) or wild-type (N) oligonucleotides. The last lane shows binding to an oligonucleotide harbouring a perfect NFkB binding site (PN). (C) Luciferase assay of reporter constructs containing SNP1 sequence corresponding to either the barred (B) or wild-type (N) allele and the empty vector pGL3-Basic. (D) Luciferase assay of reporter constructs containing SNP2 sequence corresponding to either the barred (B) or wild-type (N) allele and the empty vector pGL3-Promoter. (E) Luciferase assay of reporter constructs containing SNP1 sequence corresponding to either the barred (B) or wild-type (N) and the empty vector pGL3-Basic as well as a construct with non-ARF test-DNA. Cotransfection with 1-100 ng of E2F-1. (F) Luciferase assay of reporter constructs containing a combination of the (B) and (N) alleles in the same plasmid. The reporter construct with only the barring allele is shown as a reference, as well as the empty vector pGL3-Basic. Values on the Y-axis show the Relative Luciferase Units (RLU): Firefly reporter luciferase levels in relation to control Renilla luciferase levels.

Table S1. Oligonucleotides used in amplification, sequencing, genotyping and EMSA experiments related to Sex-linked barring in chicken.

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