Svf1 acts at the interface of the ER and the cis-Golgi. (a) Proteomic analysis of Svf1-GFP, expressed from a plasmid under the control of the endogenous promoter of Svf1 in a svf1Δ background, and mock treated WT cells is shown. Protein intensities are plotted against heavy/light SILAC ratios. Significant outliers are colored in red (P < 1⁻¹¹), orange (P < 1⁻⁴), or steel blue (P < 0.05), other proteins are shown in light blue. (b) Co-localization of GFP tagged Svf1 with mKate tagged Mnn9 (cis-Golgi) and Halo tagged Sec31 (ER, ER exit sites). (c) Evaluation of the co-localization of Svf1 dots (n = 100, triplicates) shown in d with either the cis-Golgi (Mnn9), Sec31 (ER, COPII vesicles), or both simultaneously. Scale bar = 5 µM. (d) Co-localization of GFP tagged Svf1 with Halo tagged Sec63 (ER marker) in a sec12^ts background under permissive (24°C, upper panel) and non-permissive (37°C, lower panel) temperature. Scale bar = 5 µM. (e) Co-localization of GFP tagged Svf1_G7A/G8A with Halo tagged Sec31 (ER marker) and mKate tagged Mnn9 in a sec23^ts background (upper panels) and in a sec17^ts background (lower panels) under permissive (24°C) and non-permissive (37°C) temperature. Scale bar = 5 µM. (f) Quantification of e (n = 60, triplicates).