Ssd1 localizes to P-bodies and stress granules in response to cellular stress. ssd1Δ cells (FLY2184) containing Ssd1-GFP and RFP-tagged P-body proteins Edc3, Lsm1, Dcp2, or stress granule proteins Pab1 and Pub1 were transferred to glucose-depleted medium for 10 min and analyzed by spinning disk confocal microscopy. GFP- and RFP-tagged proteins were encoded by plasmids (FLE1019 for Ssd1-GFP; see Table II for P-body and stress granule plasmids). Ssd1 colocalizes with P-body proteins in all (100%) glucose-depleted cells that have visible Ssd1 puncta (n = 75 Lsm1-RFP cells; n = 40 Edc3-RFP cells; n = 100 Dcp2-RFP cells; n = 112 Pab1-RFP cells). Furthermore, most Ssd1 puncta (∼70–90%) colocalize with P-body and stress granule proteins. Percentage of Ssd1 puncta that colocalize with P-body/stress granule proteins: 91% for Lsm1, 89% for Edc3, 73% for Dcp2, 84% for Pab1, and 88% for Pub1 (n = 10–15 cells). Parallel experiments indicate that 1 M NaCl treatment also causes Ssd1 to colocalize with P-bodies in cytoplasmic puncta (not depicted). All images represent single optical sections. Bar, 2 µm.