Figure 11

Multi-species EMSA analysis performed with recombinant Oct4 and Sox2 incubated with part of the CFTR 3' UTR. EMSA analysis [43] was performed with recombinant Oct4 and Sox2 proteins and with radiolabeled sequences showing mobility of DNA-protein complexes formed on gels. Oct4 _D refers to homodimer (Oct4/Oct4), Oct4 _H to Oct4/Sox2 heterodimer and Oct4 _M to Oct4 monomer formation. Lane 1: purified recombinant proteins Oct4 and Sox2 incubated with bovine CFTR sequence resulted in strong monomer (Oct4), weak homodimer (Oct4/Oct4), and weak heterodimer formation (Oct4/Sox2); Lane 2: purified recombinant proteins Oct4 and Sox2 incubated with mouse CFTR sequence resulted in monomer (Oct4), homodimer (Oct4/Oct4), and heterodimer formation (Oct4/Sox2); lane 3: purified recombinant proteins Oct4 and Sox2 incubated with chicken CFTR sequence resulted in no Oct4 protein binding; lane 4: purified recombinant proteins Oct4 and Sox2 incubated with sheep CFTR sequence resulted in monomer (Oct4), and heterodimer formation (Oct4/Sox2); lane 5: purified recombinant proteins Oct4 and Sox2 incubated with wallaby CFTR sequence resulted in no Oct4 protein binding; lane 6: purified recombinant proteins Oct4 and Sox2 incubated with pig CFTR sequence resulted in weak monomer (Oct4) and weak heterodimer formation (Oct4/Sox2); lane 7: purified recombinant proteins Oct4 and Sox2 incubated with platypus CFTR sequence resulted in monomer (Oct4) binding; lane 8: purified recombinant proteins Oct4 and Sox2 incubated with lemur CFTR sequence resulted in weak monomer (Oct4) and weak heterodimer formation (Oct4/Sox2); lane 9: empty lane; lane 10: control with purified recombinant protein Oct4 alone incubated with mouse CFTR sequence resulted in monomer (Oct4) and homodimer (Oct4/Oct4) binding. 27 bp EMSA oligonucleotides for each species were all derived from the genomic region orthologous to a segment containing the PORE-like sequence from mouse (ATTTGTGATGCAAAT).