A) The upper panel shows the expression of SRY and RNAPol II, respectively, in nuclear protein extracts from untransfected (line HEK293), pIRES SRY-transfected HEK-293 cells (HEK293/SRY) and 112-D cells as a negative control (Control); SRY antibody was also assayed in cytoplasmic extracts and GAPDH was used as load control in (lower panel). B) EMSA experiments showing the complex formation among labeled SRY1WT*, SRY2WT* and SRY3WT* sites (1 pmol) in the presence of 10 μg nuclear proteins from SRY transfected HEK-293 cells (lines 1, 4, 7). For the competition assay, 2 pmol of unlabeled WT oligonucleotides (SRY1WT, SRY2WT, and SRY3WT in lines 3, 6 and 9) and nonspecific oligonucleotides (T485C in lines 2, 5 and 8) were added to the reaction mixture, before the addition of labeled probes. The upper panel shows the quantification of the DNA bound in each line.