Fig. 2

Sphingolipids are required for exocytic secretion. a Bgl2 secretion in the wild-type and lcb1-100 temperature-sensitive mutant. Wild-type and lcb1-100 mutant cells were grown to early log phase in YPD liquid medium overnight at 25 °C. The cells were then shifted to 25 °C or 39 °C for 60 min. Bgl2 secretion was analyzed by western blot using anti-Bgl2 antibody. The total protein on PVDF membranes were stained with Ponceau S as loading control. b Quantification of the amounts of Bgl2 accumulation in the wild-type and lcb1-100 mutant in a. n = 3. *p < 0.01. c Invertase secretion in the wild-type and lcb1-100 mutant cells. Wild-type and lcb1-100 mutant were grown to early log phase in YPD liquid medium overnight at 25 °C. The cells were then shifted to 25 °C or 39 °C for 60 min. Then the invertase secretion were examined. d Bgl2 secretion in wild type cells treated with or without myriocin. Wild-type cells were grown to early log phase in YPD liquid medium overnight at 25 °C. Myriocin was added into the medium to a final concentration of 10 μg/ml, and the cells were incubated at 25 °C for 60 min. External and internal Bgl2 secretion was analyzed by western blot using anti-Bgl2 antibody. The total protein on PVDF membranes were stained with Ponceau S as loading control. e Quantification of the amounts of Bgl2 accumulation in a. n = 3. Three independent experiments were performed. *p < 0.01. f Invertase secretion in wild-type cells treated with or without myriocin. Wild-type yeast were grown to early log phase in YPD liquid medium overnight at 25 °C. Myriocin was added into the medium to a final concentration of 10 μg/ml, and the cells were incubated at 25 °C for 60 min. n = 3. Three independent experiments were performed. *p < 0.01