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Fig. 8. Stress granules induced by CPEB1, but not by arsenite, recruit components of dcp1 bodies. (A-D) CPEB1-induced stress granules recruit dcp1 and GW182. HeLa cells were transiently transfected with an expression vector for GFP-tagged CPEB1-
5-short. After 20 hours, cells were fixed and stained with anti-dcp1 (A-C) or anti-GW182 (D) antibodies (red). Cells were observed by confocal microscopy. As several patterns were observed, three cells are shown in A, B and C, which are representative of 70%, 10% and 20% of the stress granule-containing cells, respectively. In C, two stress granules surrounded with several dcp1 bodies have been enlarged for clearer visualization. (E) Arsenite increases the number of dcp1 bodies. HeLa/CPEB1-lg cells were induced for 16 hours with doxycycline, stressed with arsenite (as) for 30 minutes, or stressed and cultured further in the absence of arsenite for 1 hour (as +1 hour). After fixation, cells were stained with anti-dcp1 antibodies and observed by fluorescence microscopy. The graph presents the number of dcp1 bodies per cell, counted in 180 cells. (F) Recruitment of dcp1 in CPEB1-induced stress granules increases with time. HeLa cells transfected as in A were fixed at various time after transfection and stained with anti-dcp1 antibodies. CPEB1-expressing cells were counted for the presence of stress granules (SG) and the presence of dcp1 in these stress granules. Stress granule-containing cells are plotted as a percentage of CPEB1-expressing cells, whereas cells with dcp1 in stress granules are plotted as a percentage of stress granule-containing cells.
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