Figure 2.

EGFP-FKBP-LC and its associated triskelion can be reversibly clustered. (A) Clathrin-associated structures are localized to the plasma membrane and the trans-Golgi network in TRVb-EFL cells. Bar, 10 μm. (B) FK1012-A (50 nM, 2 h) leads to a clearing of cytoplasmic clathrin and the formation of large intracellular aggregates of clathrin. Clathrin puncta remain associated with the plasma membrane after cross-linking. (C) FK506 (10 μM, 2 h) reverses FK1012-A–mediated cross-linking, as clathrin distributes in a similar pattern to that of the control condition (A). (D–F) After incubation with FK1012-A, EGFP-FKBP-LC fluorescence (D) and indirect immunofluorescence against clathrin heavy chain (E) overlap almost perfectly (F).