Fig. 3. YFP-tagged EBP50 co-immunoprecipitates with myc-TRPC4 but not with the mutant lacking the last three C-terminal residues. myc-TRPC4 or myc-TRL were transiently expressed in HEK293 cells together with YFP alone or YFP-EBP50. Detergent extracts of the transfected cells were incubated with an anti-YFP antibody and protein G-coated agarose beads. 40% of the precipitated proteins were separated by SDS-PAGE, transferred to PVDF and immunoblotted with an anti-myc antibody (lower panel). Middle panel: cell lysates (17% of the input used for the co-immunoprecipitation) probed with anti-myc antibody to detect myc-TRPC4 and myc-TRL Upper panel: cell lysates probed with anti-YFP antibody to detect YFP and YFP-EBP50. Size markers indicate the molecular mass in kDa. IB, immunoblot; IP, immunoprecipitation. Results shown are representative of three independent experiments.