Figure 3

Viral protein expression upon F-actin disruption and stabilization. MDCK cells were infected with MV at an MOI of 10 (control, CD, Jaspla), or left uninfected (mock). Inhibitors were added at 12 h p.i.. At 48 h p.i., cells were surface biotinylated and subsequently lysed in RIPA buffer. ( A ) To determine the amount of MV glycoproteins expressed on the cell surface, H and F were immunprecipitated from cell lysates with specific monoclonal antibodies. Samples were then separated by SDS-PAGE, transferred to nitrocellulose and probed with AF680-conjugated streptavidin. ( B ) M protein was immunoprecipitated from cell lysates using an anti-M specific antibody. Precipitates were subjected to western blot analysis using M-specific primary antibodies and AF680-conjugated secondary antibodies. ( C ) For N, tubulin and actin staining, lysates were directly subjected to SDS-PAGE, blotted to nitrocelluose and incubated with specific primary antibodies and fluorophore-conjugated secondary antibodies. Blots were scanned with a Li-Cor Odyssey IR system.