Staphylococcal domains of Tudor-SN localize to stress granules. (A) A schematic diagram of the YFP–TSN and mC–TSN constructs; the amino acids of Tudor-SN included in each construct are shown. (B–E) HeLa cells were transiently transfected with expression vectors for (B) mC–4SN, (C) mC–SN1/2, (D) mC–SN3/4, and (E) mC–eTD. After 24 h the cells were cultured in the absence (a–c) or presence of arsenite (d–f) before processing for visualization of mC–4SN, mC–SN1/2, mC–SN3/4, or mC–eTD, respectively (red; a,d), and G3BP (green; b,e) using confocal microscopy. DAPI staining is in blue (Merge; c,f). Bar, 10 μm. (F) HeLa cells were transiently transfected with expression vectors for TSN–mC, 4SN–mC, SN1/2, and mC–eTD, and lysates prepared after 24 h. Immunoblotting was used to analyze expression. Actin was a loading control. (G) An RFP antibody was used to immunoprecipitate GFP–TSN proteins from HeLa cell lysates, and immunoblotting was subsequently used to detect interacting proteins. Pre-immune serum (PI) was used as a control.
