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Fig. 7. Effect of cycloheximide on polysomes, stress granules and mRNA degradation during heat shock. (A) Cycloheximide prevents polysome dissociation and partially restores polysomes during heat shock. Polysomes were analysed in cells incubated (from top to bottom) at: 27°C; 41°C for 60 minutes; 27°C with cycloheximide for 30 minutes; 27°C with cycloheximide for 30 minutes, followed by 60 minutes at 41°C; 41°C for 60 minutes followed by 30 minutes at 41°C with cycloheximide and; 27°C for 30 minutes with puromycin. (B) Cycloheximide prevents formation of heat-shock stress granules. Fluorescent images of cells that express PABP1-eYFP were incubated (from top to bottom) at: 27°C; at 41°C for 60 minutes; 27°C with cycloheximide for 60 minutes; 27°C with cycloheximide for 60 minutes followed by 60 minutes at 41°C; 41°C for 60 minutes followed by 60 minutes with cycloheximide and; 27°C with puromycin for 60 minutes. (C) Cycloheximide prevents the heat-shock-induced decay for PDI-2 and Tubulin. Cultures were incubated at 41°C (left lanes), 27°C with cycloheximide (middle lanes), and 27°C with cycloheximide followed by incubation at 41°C (right lanes). Analyses of the northern blot for PDI-2 and tubulin are shown below.
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