MA104 and BHK-21 cells (obtained from China Center For Type Culture Collection) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco) supplemented with 10% fetal calf serum (FCS) at 37 °C under 5% CO2. PRV (strain DN30209, isolated in a pig farm, in Heilongjiang province of China) [26 (link)] and vesicular stomatitis virus (VSV, strain Indiana, obtained from prof. Joerg Glende, University of Veterinary Medicine Hannover, Germany) were propagated on the MA104 cells and BHK-21 cells, respectively, as previously described [26 (link), 27 (link)]. In as much as previous reports showed that VSV infection on BHK-21 is not affected by depletion of cholesterol [27 (link)], we used VSV as a negative control for our studies about relationship between lipid rafts and virus infection.
Rabbit polyclonal antibodies to VP4 and VP7 were generated in our laboratory using previously described methods [26 (link), 28 ]. Anti-VSV G-protein polyclonal antibody was purchased from Abcam. An anti-β-actin monoclonal antibody was purchased from Beyotime. The secondary antibodies were purchased from BD Biosciences. Both MβCD and cholesterol were purchased from Sigma and reconstituted in DMEM and alcohol, respectively.
Rabbit polyclonal antibodies to VP4 and VP7 were generated in our laboratory using previously described methods [26 (link), 28 ]. Anti-VSV G-protein polyclonal antibody was purchased from Abcam. An anti-β-actin monoclonal antibody was purchased from Beyotime. The secondary antibodies were purchased from BD Biosciences. Both MβCD and cholesterol were purchased from Sigma and reconstituted in DMEM and alcohol, respectively.
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