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Figure 2


Fig. 2 Overproduction of Lsm4p or its C-terminus leads to aggregation in cytoplasmic foci. (A) GFP-Lsm4 (pMPSLsm4), GFP-Lsm4C (pMPSlsm4D2) and GFP-Lsm4{Delta}C (pMPSLsm4D1) were overexpressed from the MET25 promoter in BY4741 cells grown in SD-Ura-Met. Localization was examined in cells during log-phase growth. (B) Colocalization of Lsm4p aggregates with Dcp2-RFP (pRP1155) was examined in BY4741 cells grown in SD-Ura-Leu-Met during log-phase growth or 20 minutes after hypo-osmotic shock. Dcp2-RFP (pMR171) localization was examined in PGAL-LSM4 cells expressing GFP-Lsm4{Delta}C grown in SD-Ura-His-Met (to prevent competition between GFP-Lsm4{Delta}C and endogenous Lsm4p for incorporation into Lsm1-7p), 20 minutes after hypo-osmotic shock. (C) Localization of GFP-Lsm1 (pGFP-N-Lsm1), GFP-Lsm6 (pMPSLsm6), Lsm8-GFP (pMR83) and GFP (pGFP-N-FUS) was examined in log-phase cells overproducing Lsm4p (PGAL-LSM4 cells grown in SDGal-Ura) and in cells with normal levels of Lsm4p (PGAL-LSM4 cells with pUSS1 grown in SD-Ura-Met). Nuclear DNA stained with DAPI is shown in blue.





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