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Subcellular fractionation of Ist1 in different yeast strains. Yeast cell extracts were fractionated by centrifugation into soluble, cytoplasmic fractions (S) and membrane-associated pellet fractions (P). The resulting samples were analyzed for the presence of Ist1-HA (pMB241), Did2(ΔC29)-HA (pAH12), and Did2(ΔC58)-HA (pAH27) by Western blot using anti-HA antibodies (WT, SEY6210; vps4Δ, MBY3; vps4Δdid2Δ, EEY26-1; and vps4Δvps24Δ, MBY12).
