Fig. 8. T. brucei P-bodies. (A) T. brucei cells have in average 2.9 P-bodies. Projection of optical sections through puromycin-treated cells that express SCD6-eYFP. Similar Z-stacks (supplementary material Fig. S5A) were used to determine the number of P-bodies per cell: the average was 2.9 per cell (n=125) with little variation in different cell-cycle stages: 2.7 in 1K1N (n=95), 3.3 in 2K1N (n=17) and 3.3 in 2K2N (n=12) cells. (B) P-body components DHH1, SCD6 and XRNA. eYFP-DHH1 and SCD6-mChFP (left panel) or mChFP-DHH1 and XRNA-eYFP (right panel) were expressed in the same cell line from endogenous loci. Representative fluorescent images of untreated, and cells after 1 hour with puromycin, cycloheximide or heat shock (41°C) are shown. Solid arrows indicate the posterior pole of the cell. An enlargement of a heat-shocked cells expressing mChFP-DHH1 and XRNA-eYFP is shown to indicate the colocalisation of DHH1 and XRNA in all spots (dashed arrows), except at the posterior pole. (C) P-bodies and heat-shock stress granules. mChFP-DHH1 and PABP1-eYFP (left) or mChFP-SCD6 and PABP1-eYFP (right) were expressed in the same cell line from their endogenous loci. Representative fluorescent images of one untreated cell and several images of cells that have been heat shocked (1 hour 41°C) are shown. Dashed arrows point at granules that contain both a P-body marker (DHH1 or SCD6) and the stress granule marker PABP1, either colocalised or close to each other.