Fig. 8

Osh3 regulates the polarized localization of the exocyst subunit Exo70 and polarized secretion of the chitin synthase Chs3. a Exponentially growing wild type or osh3∆ cells expressing Exo70-GFP from its endogenous promoter were incubated at 32 °C for 10 min just prior to being placed on a coverslip for imaging by spinning disk confocal microscopy. Representative confocal sections showing Exo70-GFP localization in wild type and osh3∆ mutant cells are provided. Arrows point to non-polarized Exo70-GFP foci in mother cells lacking Osh3 (osh3∆). Mother (m) and daughter (d) cells are indicated. Scale bar, 2 μm. b Quantitative analysis of Exo70-GFP polarization in small-budded cells after 10 min at 32 °C. Total number of cells analyzed: wild type 10 min 32 °C n = 182, osh3∆ 10 min 32 °C n = 146. The graph shows the means and standard deviations from three independent experiments (t test, **p < 0.002). c Exponentially growing wild type or osh3∆ cells expressing Chs3-GFP from its endogenous promoter were incubated at 32 °C for 10 min just prior to imaging by spinning disk confocal microscopy. Representative confocal sections showing Chs3-GFP localization in wild type and osh3∆ mutant cells are provided. Arrows point to cortical Chs3-GFP foci mislocalized from the mother-daughter neck in cells lacking Osh3 (osh3∆). Mother (m) and daughter (d) cells are indicated. Scale bar, 2 μm. d Quantitative analysis of Chs3-GFP mother-daughter neck localization in exponentially growing cells after 10 min at 32 °C. Total number of cells analyzed: wild type 10 min 32 °C n = 75, osh3∆ 10 min 32 °C n = 144. Graph shows the means and standard deviations from three independent experiments (t test, **p < 0.002)