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Fig. 4. Expressed M-FAG remains cytoplasmic while expressed Ct/p100 causes morphological defects and induces apoptosis. (A) Schematic representation of eIF4GI outlining the individual sequences used for expression in this study. (B) HeLa cells were transfected with plasmids encoding myc-tagged isoforms of the C-terminal domains of eIF4GI. Sixteen hours after transfection, total cell lysates were prepared and proteins resolved by SDS-PAGE. The fragments were identified by immunoblotting using anti-myc antiserum; molecular mass markers are shown on the left. Owing to differences in expression levels and to aid visualisation, extracts from cells expressing C-FAG were diluted 1:10 prior to analysis. (C) Anti-myc antibody followed by goat anti-mouse IgG conjugated to FITC (green) was used to visualise the localisation of the tagged isoforms of the C-terminal domains of eIF4GI expressed in HeLa cells, as indicated. Actin was visualised with phalloidin-TRITC (red) and nuclei with DAPI (blue). Scale bars: 20 µm.
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