Fig. 4.

Functional Rab1a/b is required for HSV-1 assembly. (A) A549 cells were transfected with Rab1a/b or control siRNA and infected with HSV-1 (MOI, 5). At 18 hpi, supernatant virions were collected as described in Materials and Methods. Virus samples treated with trypsin, as well as parallel cell lysates, were separated by SDS-PAGE and blotted for pUL48 (upper blots) or tubulin (lower blots). Blots were quantified using the LI-COR Odyssey infrared imager, and the Odyssey v3.0 software was used to calculate the integrated intensity for quantification. The graph represents the amount of pUL48 in the supernatant normalized to the pUL48 signal in the cell lysate. (B) A549 cells were grown on coverslips, transfected, and infected (MOI, 3) as described in Materials and Methods. At 16 hpi cells were fixed, processed, and analyzed by transmission EM. Arrows indicate enveloped capsids, and arrowheads indicate unenveloped capsid. Bars, 500 nm.