ASFV entry is independent of clathrin-mediated endocytic pathway.

A) ASFV entry is partially inhibited by Dynasore but not by Chlorpromazine. Pretreated Vero cells with DMSO, 100 µM Dynasore (Dyn) and 20 µM Chlorpromazine (CPZ) were infected (MOI 10) for 60 min. Infected cells were analyzed by FACS and the graph shows the percentage of virus entry relative to DMSO control, measured as p72 signal. Bars represent the mean of three independent experiments (mean ±S.D., performed in duplicate). B–C) Dynamin and Clathrin are important for infection progress. Synthesis of viral proteins was measured in infected Vero cells (MOI 1) in the presence of Dyn (B) and CPZ (C) at 6 and/or 16 hpi at the indicated concentrations by Western blot with an anti-ASFV antibody. β-actin was detected as a load control. D) Viral production in the presence of Dynasore and Chlorpromazine. After 48 hpi (MOI 1) supernatants from DMSO, Dyn (100 µM) and CPZ (20 µM) treated cells were recovered. Lytic viruses were titrated in Vero monolayers and plotted in the table (n = 3). S.D., standard deviations; Dyn, Dynasore; CPZ, Chlorpromazine. *Unspecific cellular protein detected by the antibody.