Fig. 1.

Effect of clathrin depletion on the plasma membrane association of eps15R and epsin. HeLa cells were transfected with siRNA specific for the clathrin heavy chain. (A) Western blot of clathrin heavy chain-depleted HeLa cells 3 days after transfection. (B) eps15R-containing puncta at the plasma membrane after clathrin depletion. (C) Punctate epsin staining in clathrin-depleted cells. Forty-eight hours after transfection with siRNA the HeLa cells were replated on glass coverslips together with mock-transfected control cells and cultivated for another 24 h. The cells were subsequently processed for immunofluorescence by using X22 monoclonal antibody for clathrin and rabbit antisera for eps15R (26) and epsin (46). Note that the clathrin knockdown does not affect the punctate distribution of eps15R and epsin. (Scale bars: 10 μm.)