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. 2014 Jan 15;25(2):290–301. doi: 10.1091/mbc.E13-08-0448

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© 2014 van Zutphen et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — The yeast vacuole has lipase activity that depends on Atg15. Steryl ester (A), triacylglycerol (B), and free fatty acid (C) content of vacuolar fractions of wild-type, atg1, and atg15 cells grown on either rich (YPD) or autophagy-inducing (SD N⁻) media. Lipase activity in isolated lipid droplet (D) and vacuole fractions (E). Western blot (F) of proteins in crude extracts of wild-type and atg15 cells expressing either Faa4-GFP or Erg6-GFP to analyze lipid droplet autophagy or Sec63-GFP to determine ER-phagy. Cells were grown to the end of the logarithmic growth phase and shifted to SD N⁻ medium for 8 h. Single optical sections (G) of atg15-mutant cells expressing Faa4-GFP (green) and labeled with FM4-64. Cells were cultivated in SD N⁻ for 8 h, showing accumulation of GFP in the vacuole lumen. Scale bar, 5 µm. Lack of the vacuolar lipase Atg15 renders cells sensitive to the inhibitor soraphen A, which blocks de novo fatty acid synthesis (H).