Fig. 5From: Inhibitor analysis revealed that clathrin-mediated endocytosis is involed in cellular entry of type III grass carp reovirusPistop2 and CPZ inhibit GCRV104 and GCRV JX01 infection. CIK cells were treated with Pistop2 and CPZ at the indicated concentrations. a and b Rt-PCR assay of virus yield in the supernatants (infection group) and CIK cells (entry group). (unpaired t-test, * P < 0.05 and ** P < 0.01). Western blot analysis to monitor viral replication level in CIK cells. c and d Cells were pretreated with indicated inhibitors (c:CPZ 10 mM, and d: pitstop 2 5 μM) for 1 h at 4 °C, then adsorbed with virus (MOI = 50) for 30 min at 4 °C. The cultures were quickly warmed to 28 °C to start infection, and uninternalized virions were removed at 0.5 hpi. After PBS washes, cells were treated with inhibitors again for 48 h at 28 °C. The processed cells were then fixed in electron microscopy grade fixative at 4 °CBack to article page