FIG 8.

Effects of repressed expression of SEC15 on Spa2-GFP localization. Strains were grown in YPD (with or without DOX) for 3 h at 30°C, and filamentation was then induced by transferring cells to YNB–20% FCS (with or without DOX) and incubating them for 1 h at 37°C. After incubation, cells were washed twice with 1× PBS, resuspended in fresh YNB–20% FCS (no DOX), and incubated for a total of 15 min. Strains THE1-Clp10 and tetR-SEC15 that bear an allele of Spa2-GFP (polarisome) were visualized at 50 and 60 min (with or without DOX). In strain tetR-SEC15 grown with DOX, the polarisome appears mislocalized after 1 h (indicated by the white circles) and is present after 10 min of DOX removal. Fluorescent images at the tip of the growing hyphae are shown with the corresponding DIC image. Scale bar, 5 μm.