Figure 2

MaviP35 inhibits mammalian and insect cell death. (a, b) Sf9 cells were transfected with plasmids encoding MaviP35-FLAG, AcP35-FLAG or chloramphenicol acetyl transferase (CAT), then infected with an AcP35-mutant baculovirus. Untransfected cells were subjected to mock infection. (a) Survival was monitored relative to the mock-infected cells. (b) Caspase activity was detected and expressed relative to the DEVDase activity in infected cells expressing the negative control CAT protein. (c and d) Sf21 cells were co-transfected with plasmids encoding GFP and the specified proteins. After 4 h heat shock-mediated induction of transgene expression, the cells were treated with actinomycin D (c) or UV (d). Viability is expressed as the number of GFP-positive cells following exposure to apoptotic stimuli, relative to their number before treatment. (e and f) MEF (e) or LN18 glioblastoma cells (f) were co-transfected with a β-galatosidase expression plasmid and either an empty vector (solid lines), or plasmids encoding MaviP35 (dashed lines) or AcP35 (dotted lines), before treatment with the indicated doses of either cisplatin (e) or crosslinked TRAIL (f). Following 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside staining, the survival of blue (transfected) cells was scored using morphological criteria. Over 80 (e) or 150 (f) blue cells were scored for each condition per experiment. Error bars represent S.E.M. from three separate experiments (a–f)