Journal of General Virology

Fig. 4. Inhibition of apoptosis by EppoMNPV iap genes. DNA laddering assays represent total DNA extracted from 35 mm tissue culture wells, which was analysed by gel electrophoresis and ethidium bromide staining. (a) Dose–response of apoptosis inhibition in actinomycin D-induced Sf-21 cells by clone HindIII-T containing the EppoMNPV iap-2 gene. Molecular size markers are included. The dose (µg) represents the quantity of plasmid clone used to transfect 1.5x10⁶ Sf-21 cells in a 35 mm diameter tissue culture dish. Vector, pBluescriptII SK⁺. (b) The iap-2 gene is capable of preventing actinomycin D-induced apoptosis in Sf-21 cells. A description of each clone is given in the legend to Fig. 1(a). A 2 µg aliquot of each clone was used for each transfection assay. Cellfectin, transfection control. (c) IAP-1 causes a delay in actinomycin D-induced apoptosis in Sf-21 cells. A description of each clone is given in the legend to Fig. 1. 18h and 30h, Incubation of Sf-21 cells with actinomycin D for 18 h or 30 h respectively prior to analysis for oligonucleosomal laddering; Cellfectin, transfection control. (d, e) IAP-3 and IAP-4 do not prevent actinomycin D-induced apoptosis in Sf-21 cells. The clones tested are described in the legend to Fig. 1(c) and (d). Cellfectin, transfection control. (f) Cell death (%) as determined by trypan blue vital staining and microscopic examination. ActD, apoptosis induced by actinomycin D.

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