Extended Data Fig. 1: Golgi localization of various proteins is not affected under hypo-osmotic or ethanol treatment conditions and in glycerol-deficient cells.
From: Glycerol mediates crosstalk between metabolism and trafficking through the golgin Imh1
(a) Golgi-resident proteins are not affected under hypo-osmotic or ethanol treatment conditions. Sec7-mRFP (late-Golgi marker), Arf1-mRFP (Golgi marker), Vps53-GFP, or GFP-Ypt6 were expressed in WT cells. The live cells were cultured in YPD medium until mid-log phase. For hypo-osmotic shock treatment, the medium was removed, and the cells were suspended in water for 5 min. For ethanol treatment, the medium was removed and the cells were suspended in YP medium containing 3% ethanol for 10 min. Scale bar, 5 μm. (b) Golgi-resident proteins are not affected in glycerol-deficient cells. Sec7-mRFP (late-Golgi marker), Arf1-mRFP (Golgi marker), Vps53-GFP, or GFP-Ypt6 was expressed in WT, gpd1∆, and gpp1/2∆ cells. (c) The localization of Golgin protein Rud3 and Sgm1 is not affected in glycerol-deficient cells. mCH-Rud3 or Sgm1 was expressed in WT and gpd1∆ cells. (b-c) Live cells were observed in mid-log phase. Scale bar, 5 μm. (a-b) The number of Sec7-mRFP, Arf1-mRFP, Vps53-GFP, and GFP-Ypt6 puncta was determined using ImageJ Fiji software (N = 3, n = 50). Data are presented as the mean ± SD of three independent experiments. (N.S., not significant; one-way ANOVA with Dunnett’s post hoc multiple comparison test, no treatment or WT cells were used as reference) (c) The number of mCH-Rud3 and Sgm1 puncta was determined using ImageJ Fiji software (N = 3, n = 50). Data are presented as the mean ± SD of three independent experiments. (N.S., not significant; two-sided unpaired t test).
