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. 2019 Jun 6;294(29):11323–11332. doi: 10.1074/jbc.RA119.008591

Figure 5.

Figure 5.

Phosphorylation of Exo84 negatively regulates exocytic secretion in cdc34-2 mutant. A, the phosphodeficient exo84-A mutant rescued the Bgl2 secretion defect of the cdc34-2 cells. The cdc34-2 and cdc34-2 exo84-A mutants were grown at 25 °C and then shifted to 37 °C for 1.5 h. Internal (In) and external (Ex) pools of Bgl2 were examined by Western blot analysis. Alcohol dehydrogenase (Adh1) was probed as a protein loading control. B, quantification of Bgl2 accumulation in the cdc34-2 and cdc34-2 exo84-A mutants at 37 °C. Error bars represent S.D. *, p < 0.01; n = 3. C, cells were fixed with permanganate and processed for thin-section EM. The cdc34-2 cells accumulated a large amount of post-Golgi secretory vesicles at the restrictive temperature, whereas cdc34-2 exo84-A double mutant did not. SV and the arrow indicate one of the vesicles. Scale bar, 2.5 μm. D, quantification of accumulated vesicles in the cdc34-2 and cdc34-2 exo84-A mutants at 37 °C. Error bars represent S.E. *, p < 0.01; n = 15. AU, arbitrary unit.