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. 2006 Jan 2;172(1):55–66. doi: 10.1083/jcb.200510016

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Figure 2. — Sro7-HA ₃ p from yeast extracts binds preferentially to GTPγS-Sec4p. (A) Expression of SRO7-HA ₃ suppresses the cold sensitivity of an sro7Δ sro77Δ strain. Wild-type (WT), sro7Δ sro77Δ, and SRO7-HA ₃ sro77Δ yeast strains were spotted in 10-fold dilutions onto YPD media plates and grown at the indicated temperatures. (B) Extracts of a wild-type or SRO7-HA ₃ strain (HA) were subjected to Western blot analysis using an α-HA antibody. Ponceau staining is shown as a loading control. (C) Sro7-HA₃p binds preferentially to GTPγS-Sec4p. GST-Sec4 (Sec4), GST-Ypt1 (Ypt1), or GST immobilized on glutathione beads were incubated with an extract of an SRO7-HA ₃ strain, and copurifying proteins were subjected to Western blot analysis using the indicated antibodies. GST-Sec4p and GST-Ypt1p were either stripped of nucleotide (NF) or loaded with GTPγS (GTP) or GDP. Ponceau staining of the Western blot is shown as a loading control. The input lanes represent 0.2 and 0.3%, respectively.