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. 2012 Oct 25;7(10):e48189. doi: 10.1371/journal.pone.0048189

Figure 4. Interaction of Sp transcription factors with the MDR1 promoter.

Figure 4

(A) EMSA with probe-C in lanes 1–9, and hCMEC/D3 nuclear protein in lanes 2–9. Protein-binding was blocked with a consensus Sp oligonucleotide, a mutated Sp oligonucleotide (Sp-mut.) and a consensus AP2 oligonuceotide (lanes 3–5). Supershifts were attempted with antibodies to Sp3, Sp1, TFIID and NFY (lanes 6–9). The nuclear protein produced five shifted bands, of which three (b,c,d) were reduced by anti-Sp3. (B) EMSA with probe-C in lanes 1–7 and Caco-2 nuclear protein in lanes 2–7. The nuclear proteins produced 4 major shifted bands (f–i). Protein-binding was blocked with consensus oligonucleotides for Sp or AP2 (lanes 3 & 4). Supershifts with specific antibodies (lanes 5–7) indicated that band ‘f’ was produced by Sp1, bands ‘g’ and ‘h’ by Sp3. Band ‘i’ was substantially reduced by all treatments.