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Fig. 3 | Virology Journal

Fig. 3

From: Entry of sapelovirus into IPEC-J2 cells is dependent on caveolae-mediated endocytosis

Fig. 3

Caveola/raft-dependent endocytosis is required for PSV infection. a Cholera toxin B (CTB) uptake was blocked by nystatin or MβCD. PK-15 cells were mock treated or pretreated with nystatin or MβCD at indicated concentrations for 1 h, and then 10 μg/ml Alexa Fluor 594 conjugated-CTB was added and incubated at 37 °C for 60 min before observed using confocal microscopy. b-c Nystatin and MβCD reduced PSV infection. IPEC-J2 cells were pretreated with increasing concentrations of nystatin or MβCD for 1 h at 37 °C, and then cells were infected with PSV (MOI = 0.5). At 24 hpi, cells were lysed and virus titers were determined by TCID50 or viral VP1 expression levels were detected by western blot. The horizontal line shows the subtoxic concentrations of nystatin or MβCD on IPEC-J2 cells. d The effect of siRNA against caveolin-1 was determined by RT-qPCR. e-f Cells were transfected with siRNA against negative control (siNC), siRNA against caveolin-1 (siCav), or cells were transfected with EGFP, EGFP-tagged wild-type caveolin-1 (Cav-WT), caveolin 1 dominant negative mutant (Y14F) (Cav-DN). At 24 h after transfection, cells were infected with PSV, and virus VP1 protein levels were detected by 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

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