Figure 5

Functional analysis CNS87 mutants. A) Schematic representation of the footprints position on the CNS87 sequence. Three different CNS87 mutants were generated and cloned into the TK-pGL3basic vector as described for the wild-type sequence; B) All the CNS87 mutants were transiently transfected in PC Cl3 thyroid cells. At 48 h post-transfection, transcriptional activity was determined as the firefly over renilla luciferase activity. Data are expressed as fold induction over the transcription obtained with TK-pGL3basic, whose value was set at 1.0. Data are means ± SD from 3-4 independent experiments, each performed in duplicate; C) HeLa cells were transiently transfected with the CNS87-TK reporter construct in the absence or in the presence of increasing concentration of the expression vector encoding TTF-1. Folds of activation are considered as ratio between values obtained with and without co-transfection of the expression vector and the transcriptional activity was determined as the firefly over renilla luciferase activity. Data are means ± SD from 3-4 independent experiments, each performed in duplicate.