Fig. 6. Electrophoretic mobility shift assay demonstrating that VegT binds Tbs1. A ³²P-labelled 36 base pair probe including Tbs1 was incubated with uncharged reticulocyte lysate (lanes 1-3) or with in vitro translated HA-tagged VegT (lanes 4-9). Assays in lanes 2, 5 and 8 included an excess of unlabelled probe as competitor (+) and assays in lanes 3, 6 and 9 included an excess of unlabelled mutated probe (M). Supershift assays in lanes 7-9 included a rat anti-HA monoclonal antibody, with incubation carried out for 10 minutes at 4°C. A specific shift is visible in lanes 4 and 6, and these are ‘supershifted’ in lanes 7 and 9.