Figure 3

Coimmunoprecipitation of distinct epitope-tagged TSC1 isoforms. TSC1myc and TSC1V5 were co-expressed with TSC2 in HEK293T cells. The distinct tagged TSC1 isoforms were immunoprecipitated and the presence of other coimmunoprecipitated tagged isoforms was investigated by immunoblotting. (A) Coimmunoprecipitation of TSC2 and TSC1myc with TSC1V5. TSC1V5 was co-expressed with TSC2 and TSC1myc. TSC1V5 was immunoprecipitated with anti-V5 affinity beads, washed and the immunoprecipitates analysed by immunoblotting. TSC1myc and TSC2 were detected in the TSC1V5 immunoprecipitate. (B) Coimmunoprecipitation of TSC2 and TSC1V5 with TSC1myc. TSC1myc was co-expressed with TSC2 and TSC1V5 or V5-tagged β-lactamase (lacV5). TSC1myc was immunoprecipitated with anti-myc affinity beads, washed and the immunoprecipitates analysed by immunoblotting. TSC1V5 and TSC2, but not lacV5, were detected in the TSC1myc immunoprecipitate. As an extra control, TSC2 and TSC1V5 were co-expressed with myc-tagged β-lactamase (lacmyc), and immunoprecipitated with anti-myc affinity beads. TSC1V5 was not detected in the lacmyc immunoprecipitate.