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Figure 3 | Virology Journal

Figure 3

From: Egress of HSV-1 capsid requires the interaction of VP26 and a cellular tetraspanin membrane protein

Figure 3

Western blot of CTMP-7 molecules in purified HSV-1 viral particles. a. Immunoblot showing the distribution of cellular CTMP-7 protein in HSV-1 infection. KMB-17 cells infected with HSV-1 (1 MOI) or uninfected were collected at 48 h post-infection and centrifuged at 10,000 rpm for 10 min at 4°C to separate debris and supernatant. The supernatant from infected cells was concentrated to 1/50 by centrifugation at 40,000 rpm for 4 hours at 4°C, re-suspended, and separated via a sucrose density gradient centrifugation. The fraction with highest titer was used in immunoblot. 1: Control cells debris; 2: Control cells supernatant; 3: The supernatant from infected cells; 4: The pellet from infected cells; 5: Purified virions from sucrose gradient centrifugation. The antibody in this immunoblot is mouse polyclonal antiserum against CTMP-7. b. Western blot analysis with anti-HSV1 antibody. The purified virus virion was electrophoresed by 10% SDS-PAGE, transferred onto nitrocellulose membranes, and used for Western blotting analysis with anti-HSV-1 antisera specific for the virus protein at 1:500 dilution. Signals were detected by an ECL system (Pierce). 1: Control cells debris; 2: Purified virions from sucrose gradient centrifugation; 3: The supernatant from infected cells; 4: The pellet from infected cells.

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