FIGURE 5.
GST pulldown analysis of the TSC1-TBC1D7 interaction. a and b, in vitro GST pulldown using purified proteins. SDS-PAGE gels were stained with Coomassie Brilliant Blue. Three independent GST pulldown experiments were performed, and all results were highly reproducible. a, wild-type GST-TBC1D7(1–293) pulls down untagged wild-type TSC1(939–992) and TSC1(939–992) mutants. b, wild-type GST-TBC1D7(1–293) and GST-TBC1D7(1–293) mutants pull down untagged wild-type TSC1(939–992). c and d, semi-quantification of the in vitro GST pulldown analysis. Purified wt/mutant TSC1 and TBC1D7 proteins were used for the pulldown analysis and the SDS-PAGE gels were stained using Oriole Fluorescent Gel Stain (Bio-Rad). A plot of relative densitometry values of the TSC1 is shown. Error bars indicate the mean ± 1 S.D. from three separate experiments.