Fig. 1: Glycerol addition reverses defective Imh1 localization induced by hypo-osmotic shock.
From: Glycerol mediates crosstalk between metabolism and trafficking through the golgin Imh1
a, Glycerol rescues Imh1 mislocalization in wsc1∆ cells. mCH–Imh1 was expressed in WT and wsc1∆ cells. Cells were grown to midlog phase in YPD medium, the medium was removed and the cells were suspended in YP medium with 0.1 M glycerol, NaCl or sorbitol. b, mCH–Imh1 is mislocalized from the late Golgi while Sec7–GFP remains in place under hypo-osmotic shock. mCH–Imh1 was coexpressed with Sec7–GFP in WT cells. Cells were grown to midlog phase in YPD medium. For hypo-osmotic shock treatment, the medium was removed and the cells were suspended in water (with different dilutions of medium as indicated in the figure; 100% indicates a suspension of cells in water) for 5 min. The detailed procedures are described in Methods. c, Mislocalized mCH–Imh1 after hypo-osmotic shock treatment is restored by the addition of glycerol-supplemented medium. mCH–Imh1 was coexpressed with Sec7–GFP in WT cells. Cells were grown to midlog phase in YPD medium. For the hypo-osmotic shock treatment, the medium was removed and the cells were suspended in water for 5 min. Then, the cells were suspended in YP or in YP containing 0.1 M glycerol, NaCl, sorbitol or maltose. The ratio of colocalization between Imh1 and Golgi marker Sec7 was quantified (N = 3, n = 50). PCC, Pearson’s correlation coefficient. Data are presented as mean ± s.d. of three independent experiments. NS, not significant. Statistical analysis was conducted using a one-way ANOVA with Dunnett’s post hoc multiple-comparison test. The WT strain without treatment was used as a reference. Scale bars, 5 μm.
