Fig. 6

Binding of T. cruzi nuclear proteins to the Pr77, DPE-mutated Pr77, and DPE-bearing sequences. a Double-stranded DNA corresponding to the Pr77 (dsPr77), or Pr77 sequence mutated at the DPE motif (dsPr77M1), were 5′ end radiolabelled with γ-ATP³² and loaded onto native 6 % polyacrylamide gels with (+) or without (−) incubation with T. cruzi nuclear proteins (NP). Competition assays were performed by adding as competitors (C) non-labelled DNAs corresponding to the Pr77 sequence (dsPr77), a mixture of aptamers (dsApt) or dsPr77M1 to the reactions at a ratio of 1:50 or 1:5 as indicated in each panel. b Double-stranded DNA corresponding to the Pr77 sequence bearing the DPE motif (dsDPE, nucleotides 12 to 33 of the Pr77 sequence) was 5′ end radiolabelled with γ-ATP³² and loaded onto native 12 % polyacrylamide gels with incubation (+) or without (−) incubation with T. cruzi nuclear proteins (NP). Competition assays were performed by adding as competitor (C) non-labelled dsDPE DNA to the reactions at a ratio of 1:5. Shifted bands are indicated with an arrowhead, and the free probe indicated as ‘FP’