Figure 8.

Six3 interaction with Crygf promoter DNA. (a) In vitro expression of Six3. Six3-pcDNA3.1 was transcribed and translated with reticulocyte lysate containing [³⁵S]methionine. The correct 37 kDa band for Six3 is apparent (lanes 1 and 2). As a control in vitro expression was performed with the pcDNA3.1 plasmid without the cDNA sequence coding for Six3 (lane 4). (b) Affinity binding of Six3 to Crygf promoter sequences. The immobilised Crygf promoter sequences (–184–145) and (–164/–115) specifically precipitate Six3 (lane 1). As a control Six3 was bound to the immobilised (–226/+46) Crygf fragment. The free DNA sequences (–214/–165), (–184/–145) and (–164/–115) displace Six3 from the immobilised fraction (lane 2) into the supernatant (lane 3). Therefore, Six3 interacts with the Crygf promoter between nucleotides –214 and –115.