Figure 5.

Clathrin modulates Ebola infectious virus-like particle (EBOV-iVLP) entry into HeLa cells. A, Downregulation of clathrin heavy chain (CHC) in HeLa cells by small interfering RNA (siRNA), as shown by Western blotting. B, Clathrin-depleted HeLa cells and control cells were infected with EBOV–iVLPs that mediate expression of Renilla luciferase, which was used as readout for infectivity. Despite a 75% downregulated CHC, the reduction of EBOV-iVLP infection was only 25%. C and D, Virus-like particle (VLP) adsorption was performed for 1 hour at 4°C followed by a 15-minute temperature shift to 37°C. Colocalization of VLPs (stained with anti-EBOV-VP40 antibody) with either clathrin or early endosome antigen 1 (EEA1), and transferrin with clathrin or EEA1 was determined by immunofluorescence analysis. G, Quantification of VLPs colocalized with clathrin, EEA1, or caveolin-1 at indicated time points. Data are based on 3 independent experiments. Error bars represent standard errors. Scale bar: 5 μm.