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. 2014 May 20;289(28):19306–19316. doi: 10.1074/jbc.M114.556944

FIGURE 4.

FIGURE 4.

Single and double amino acid exchanges within the C-terminal region affect stability and activity of Tgl3p. A, amino acid sequence of the C-terminal 5 kDa of Tgl3p is shown, and positions of single and double amino acid exchanges are marked with arrows. All variants were N-terminal HA-tagged. B, homogenates (Hom), 30,000 × g (30g) microsomes, 40,000 × g (40g) microsomes, and LD) from strains harboring full-length Tgl3p, Tgl3p variants with single amino acid exchanges F630A and L632A, and Tgl3p variants bearing double amino acid exchanges D633A/D634A and I635A/I636A were analyzed by Western blotting. Primary antibodies were directed against the HA tag, Wbp1p (ER marker), and Ayr1p (LD marker). 10 μg of protein were loaded per lane. C–E, the relative amounts of TG from the respective strains were analyzed from cells grown to the stationary phase. Results are the average from at least three independent experiments with S.D. values (error bars) as indicated.