Figure 1.

Purified Virions Are Infectious

(A) Vero cell lysate and 30 μl of growth media were analyzed by western blotting to determine the presence of VP1 capsid protein and GAPDH expression.

(B) The lysate from RPTE cells infected by gradient purified virions was separated by SDS-PAGE and analyzed by western blotting for the VP1 and VP2/VP3 capsid proteins.

(C) RPTE cells were grown on glass coverslips and infected with virions; 3-dpi slides were fixed with 4% paraformaldehyde and stained with antibodies against VP1 (green) and VP2/VP3 (red). The nuclei were visualized with DAPI (blue). Scale bars, 10 μm.

(D) Negative-stain electron micrograph of virions present in the Vero cell lysate.

(E) Partially purified virions following centrifugation through a cesium chloride gradient.

(F) Purified virions following final purification step through a centrifugal concentrator (right).

(G) Purified virions suspended in a layer of vitreous ice following cryo-grid preparation.

Scale bars, 100 nm.