Figure 6.

INO1 expression in wild-type, OPI1^ffat, and scs2Δ strains and localization of Opi1p-GFP in wild-type or Opi1^ffatp-GFP in scs2Δ and OPI1^ffat strains, respectively, after a shift to I⁻C⁺ medium. A, overnight cultures of YCY3 (wild type), YCY5 (OPI1^ffat), and YCY7 (scs2Δ) expressing genomic Opi1p-GFP or Opi1p^ffat-GFP, respectively, were diluted to A₆₀₀ = 0.2 in I⁺C⁺ medium and allowed to grow to mid-logarithmic phase at 30 °C. Cells were harvested by centrifugation, washed and resuspended in I⁻C⁺ medium, and incubated for 3 h. Inositol was added back after 3 h of inositol starvation. Samples were taken at 0, 1, 2, and 3 h of inositol starvation and 30 min after adding back inositol. Total RNA was isolated and analyzed by RT-PCR as described under “Experimental procedures.” Solid triangles, wild type; solid squares, OPI1^ffat; solid crosses, scs2Δ. B, Opi1p-GFP localization in wild type and Opi1p^ffat-GFP in OPI1^ffat at 0 and 2 h after a shift to I⁻C⁺ medium. Cells were imaged by fluorescence microscopy. A representative z-section was chosen for each image. Scale bar, 5 μm.