Fig. 2.

A, In vitro effects of the PAX8 promoter mutation. Both PAX8 gene promoter constructs (PAX8proWT and PAX8proMUT) were transiently transfected into HeLa cells and into PCCL3 cells. A, Using 1 μg of plasmid DNA of each promoter construct in both cell lines, a significantly reduced luciferase expression was observed with the PAX8proMut (the red and white hatched bar shows transfection in PCCL3 cells, the red and black hatched bar represents the results in Hela cells) compared with the PAX8proWT (black bar). The empty vector is shown as a white and black hatched bar. B, EMSA with nuclear extracts from PCCL3 with PAX8proWT and PAX8proMUT. Nuclear extracts isolated from PCCL3 cells were incubated with oligonucleotides containing either the WT or the mutant PAX8 promoter sequences. A specific shift with the WT promoter sequence is observed. A specific binding activity is observed only with the WT PAX8 promoter sequence (green arrow).