Fig. 4

Interaction of Cryptic promoter region with endogenous Snail in nuclear lysates of PANC1 cells. Total nuclear protein extract (NPE) was isolated from PANC1 cells that express Snail endogenously. (a) Schematic of oligonucleotide duplex corresponding to Cryptic promoter region 15 bp upstream and 15 bp downstream of the Snail binding element was used for electrophoteric shift, (b) Schematic of the Snail binding element was mutated (showed in red). (c) the schematic complex of oligonucleotide supershift is depicted. (d) Lane 1 represents the biotinylated probe. Lanes 2 and 3 represent the incubation of increasing amounts of NPE with wild type probe. Lanes 4 and 5 are obtained upon incubating the NPE with the wild-type oligonucleotides with IgG control or Snail specific antibodies. Lanes 6 represents the mutated Snail binding element Lane 7 represents the incubation of the NPE with SBE mutated oligonucleotide. NPE: nuclear protein extract, * represents 10 μg NPE; blue and red arrows represent shift and supershifts, respectively