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. 2011 Jan 15;22(2):216–229. doi: 10.1091/mbc.E10-04-0317

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© 2011 Lorente-Rodríguez 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 8: — Depletion of PI(4)P in vivo causes a block in ER-to-Golgi transport and influences Golgi morphology. (A) Wild-type (SEY6210) and pik1–83 (AAY104) strains grown to a mid-log stage at 26°C were shifted to 37°C for 45 min, pulsed for 7 min at 37°C, and chased at 37°C for the indicated times. The ER (p1), Golgi (p2), and vacuolar (m; mature) forms of CPY are indicated. Note the block in maturation of p1 CPY in the mutant. (B–E) Wild-type (CBY2970, CBY2973, CBY2974) and pik1–83 (CBY2980, CBY2983, and CBY2984) cells expressing Sec63p-GFP (C), Sec21p-GFP (D), or Sec7p-GFP (E) were grown on YPD at 26°C to 0.4 OD₆₀₀/ml and shifted to 37°C. (B) Cultures were analyzed for growth over time by absorbance. (C-E) pik1–83 cells retain ER (C), cis-Golgi (D), and late-Golgi (E) structures. Scale bars: 5 μm. A quantitative analysis of ER and Golgi morphologies observed in a population of cells at the 1-h time point is provided in Supplemental Figure S5.