FIGURE 3.

Mutations of the clathrin box region alter interaction of Gga2 with clathrin in cell lysates. a, effect of Gga2 mutations on clathrin binding to Gga2 and Ent5 in lysates. The indicated mutations were generated at the endogenous Gga2 locus. Clathrin was immunoprecipitated from indicated strains with a polyclonal clathrin light chain antibody, and interacting Gga2 and Ent5 were detected by immunoblot analysis. The left panel shows representative immunoprecipitation (I.P.) results. The right panels show intensity analysis as described in Fig. 1. b, effect of Gga2 mutations on Gga2 binding to clathrin and Ent5 in lysates Indicated mutations were generated at the endogenous Gga2 locus and GFP-tagged at the C terminus. Gga2-GFP alleles were immunoprecipitated with a monoclonal GFP antibody, and interacting clathrin and Ent5 were detected by immunoblot analysis.