Analysis of Svf1 acetylation and analyses of Svf1 mutants. (a) Mass spectrometric analysis of the N-terminus of Svf1 showing that it is N-terminally acetylated. MS/MS spectrum extracted from MaxQuant data from a GFP pulldown of Svf1. Detected y-ions are labeled in red, b-ions are labeled in blue. The sequence showing all identified y- and b-ions is shown in the top right position. (b) Tetrad analysis of the svf1Δpl.Grh1_1-18aaSvf1_19-481-GFP (blue and green, respectively) mutants crossed with sur2Δ (red). (c) Evaluation of the binding of the AHs from WT, V12D mutant, and G7A/G8A mutant to the ER expressed as cytosol/ER ratio in %. Intensities of the GFP signal measured in cytosolic areas were divided by the intensities of the GFP signal at the ER measured in 0.065 µm². The signal of Sec63-Halo was used to differentiate between Cytosol and ER areas. n ≤ 48.