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Fig. 7. Stress granule assembly and dcp1 bodies depend on the functionality of the mRNA binding domain. (A) The four CPEB1 isoforms can assemble stress granules. HeLa cells were transiently transfected with expression vectors for the indicated GFP-tagged CPEB1 isoforms. After 24 hours, cells were fixed, stained with anti-eIF3 antibodies (red) and observed by confocal microscopy. The figure illustrates only cells harbouring stress granules. (B) All CPEB1 isoforms can colocalize with dcp1 bodies. Cells obtained as in A were stained with anti-dcp1 (red) and observed by confocal microscopy. The figure illustrates only cells harbouring small CPEB1 foci. (C) A C-terminal truncation of CPEB1 abrogates the colocalization with dcp1 bodies. HeLa cells were transiently transfected with an expression vector for an RRM- and Zn finger-deleted CPEB1-long. After 24 hours, cells were fixed and stained with anti-dcp1 antibodies (red). (D) A C-terminal truncation of CPEB1 abrogates the localization in stress granules. HeLa cells transfected as in C were stressed with arsenite for 30 minutes, fixed and stained with anti-eIF3 antibodies (red). (E) H545A and F314A point mutations abrogate the localization of CPEB1 in dcp1 bodies. HeLa cells were transfected with an expression vector for GFP-tagged CPEB1-long-H545A or -F314A and stressed as in D. Cells were stained with anti-dcp1 antibodies (red).
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