Fig. 2.

PSV entry and infection are independent of clathrin-mediated endocytic pathway. a The effect of CPZ (10 μM) on transferrin uptake in PK-15 cells was observed using confocal microscopy. b-c Viral yield assays in CPZ or mock-treated IPEC-J2 cells. Cells were pretreated with CPZ at the subtoxic concentrations (b, horizontal line) and then infected with PSV in the presence of CPZ for 1 h. At 24 hpi, cells were collected and the progeny virus titer was determined by TCID₅₀ and the expression level of viral VP1 proteins was analyzed by western blot. d The effect of siRNA against clathrin heavy chains (CHC) was determined by RT-qPCR. e-f IPEC-J2 cells were transfected with siRNA against negative control (siNC), or two siRNAs targeting CHC or cells were transfected with EGFP, EGFP-tagged wild-type Eps15 (Eps15wt), or DN mutant Eps15 (EpsΔ95/295). At indicated time after transfection, cells were infected with PSV and processed for western blot. Data are presented as means ± SD. The error bars indicate the standard deviations of triplicate samples from one of two independent experiments. Bars, 30 μm. ^*P < 0.05; ^**P < 0.01