Figure 4. Scd6 localizes to and affects granules.

A) Strain bearing chromosomal GFP-Edc3 was transformed with plasmid expressing Pbp1-mCherry. Cells were grown to 0.3–0.4 OD₆₀₀ in appropriate minimal media with 2% glucose and then stressed in minimal medium lacking glucose for 10 minutes before observing under the microscope. B) Strain bearing chromosomal GFP-Scd6 was transformed with plasmid expressing Edc3-mCherry. Cells were grown to 0.3–0.4 OD₆₀₀ in appropriate minimal media with 2% glucose and then stressed in minimal medium lacking glucose for 10 minutes before observing under the microscope. C) Strain bearing chromosomal GFP-Scd6 was transformed with Pbp1-mCherry. Cells were grown to 0.3–0.4 OD₆₀₀ in appropriate minimal media with 2% glucose and then stressed in minimal medium lacking glucose for 10 minutes before observing under the microscope. D) Wild type or scd6Δ strains were transformed with vector expressing Dcp2-mCherry. Cells were grown and stressed in absence of glucose as described in A and observed under the microscope. E) Wild type or scd6Δ strains were transformed with vector expressing Dcp2-mCherry and Pab1-GFP. Cells were grown up to 0.3–0.4 OD₆₀₀ stressed by adding 0.5% sodium azide (or equal of water for control cells) to the medium, incubated for 30 minutes at 30°C before observing under microscope. F) Wild type strains expressing Edc3-mCherry and Pab1-GFP were transformed with either empty vector or vector driving over-expression of SCD6 or SCD6ΔRGG from the Gal-inducible promoter. These strains were grown to 0.3–0.4 OD₆₀₀ in appropriate minimal media with 2% sucrose as carbon source followed by growth in medium containing 1.8% galactose-0.2% sucrose for 2 hours. Cells were then observed under the microscope. G) eIF4G-GFP, eIF4E-GFP and eIF3b-GFP strain were transformed with empty vector or vector driving over expressing SCD6 under Gal-inducible promoter. These strains were grown to 0.3–0.4 OD₆₀₀ in appropriate minimal media with 2% sucrose as carbon source followed by growth in medium containing 1.8% galactose-0.2% sucrose for 2 hours. Cells were then observed under the microscope.