Effect of HSV1 infection on APP processing in SHSY5Y cells. (A) SHSY5Y human neuroblastoma cells were either untreated (cont), infected with herpes simplex virus type 1 (HSV1), or treated with the protein synthesis inhibitor cycloheximide (CH), then incubated for a further 6 h. Cell lysates were subjected to Western blot analysis, using an anti-C-terminal APP antibody (Sigma Aldrich A8717) at 1:4000 dilution. Each lane contains cell lysate prepared from a single flask (two flasks were used per treatment). Several full-length APP bands are clearly visible. Arrows indicate the band intensified by HSV1 infection. (B) Quantification of 55 kDa band in control (cont), cycloheximide-treated (CH), or HSV1 infected cells, assessed using Syngene GeneTools software. Values show average band height from five independent experiments for cells treated for 6 h, each involving two or four separately processed flasks. Bars show standard deviation. Treatments resulting in values significantly different from control are marked * (indicates p < 0.002; ANOVA). (C) Effect of time period after inoculation with virus on height of 55 kDa band. SHSY5Y human neuroblastoma cells were either untreated (cont), infected with HSV1, or treated with cycloheximide (CH), then incubated for a further 3, 6 or 9 h. Cell lysates were analysed by Western blotting as in Fig 1A. Arrows indicate the band intensified by HSV1 infection.
Shipley et al. BMC Microbiology 2005 5:48 doi:10.1186/1471-2180-5-48