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. 2000 Mar 15;28(6):1473–1480. doi: 10.1093/nar/28.6.1473

Figure 3.

Figure 3

DNA binding and trans-activation by SOX8. (A) DNA binding. Lanes 1–8 contain equal amounts of full-length mouse SOX8, lanes 9 and 10 contain equal amounts of in vitro transcription/translation product of an antisense Sox8 construct (SOX8-AS). Lanes 1–8 all contain 2 ng of labelled oligonucleotide. Unlabelled non-specific competitor DNAs were included in lanes 3 and 4, as shown. Lanes 5 and 6 contain a 100-fold excess of unlabelled competitor oligonucleotide. Lanes 7 and 8 contain equimolar and twice molar amounts, respectively, of unlabelled oligonucleotide B. Oligonucleotide A contains the SoCM consensus sequence AACAAAG and B contains the SryC sequence AACAAT (21). The position of the SOX8 protein–DNA complex is indicated on the left. (B) GAL4 trans-activation assay. COS1 cells were transfected with the GAL4 fusion constructs shown on the left. The numbers represent mouse SOX8 amino acid intervals fused with GAL4. The right panel shows luciferase activity produced by each GAL4–SOX8 construct, relative to GAL0 = 1, with standard errors calculated from four replicate transfections. Similar results were observed in three independent experiments, each involving triplicate or quadruplicate determinations.