Fig. 4

Deletion of the Gic1 and Gic2 in sec3 mutant cells defects in exocytosis. a sec3∆Ngic1∆gic2∆ triple mutant cells have Bgl2 secretion defects at 25 °C, with a significantly greater defect at 37 °C. Single, double, and triple mutants, as well as sec10-2 mutants that are known to have Bgl2 secretion defects at 37 °C, were grown in YPD at 25 °C to early log phase and then grown at 25 °C or shifted to 37 °C for 90 min. The internal and external pools of Bgl2 are detected by Western blot using anti-Bgl2 antibody, and total protein on the PVDF membrane stained with Ponceau S was used as a loading control. b Quantification of the amounts of Bgl2 accumulation in cytosol in (a). Error bars represent standard deviation. n = 3. c sec3∆Ngic1∆gic2∆ triple mutant cells have invertase secretion defects. Single, double, and triple mutants, as well as sec10-2 mutants were grown in YPD at 25 °C to early log phase and then grown at 25 °C or shifted to 37 °C for 90 min and the invertase secretion of each strain was measured. *p < 0.01. Error bars represent standard deviation. n = 3. d Triple mutants accumulated a large number of vesicles. Single, double and triple mutants were grown in YPD at 25 °C to early log phase. Cells were fixed and imaged using an electron microscope. e Quantification of images in (d). 10 cells were counted per group, bars represent standard error. *p < 0.05, student’s t-test