Figure 4.

Design of minicircles containing one and two κB binding sites efficiently bound by NF-κB p50/p50. (A and D) Sequence of 95 bp minicircles containing one and two κB binding sequences as indicated by the rectangular frame. (B and E) EMSA showing titration of minicircle by increased concentrations of NF-κB p50/p50 as indicated under Materials and Methods. (C and F) Plot of data from panels B and E, respectively; data were fitted to a square hyperbola using non-linear least squares analysis and according to previously reported equations (20) related to unbound (▴), singly bound (♦) and doubly bound (▪) DNA.