Prolonged VPS8D hairpin induction leads to cytoplasmic dispersal of Dop1p, Vma4p and Scr7p.
A: Severe mislocalization of Dop1p upon VPS8D knockdown. Growing cells expressing DOP1 -mNeon were fixed and the nuclei stained with DAPI. DIC and fluorescent images of the wild type and VPS8Dknockdown cells are shown in the left and right panels, respectively.
B: Severe mislocalization of Vma4p upon VPS8D knockdown. Growing cells expressing VMA4 -mNeon were fixed and the nuclei stained with DAPI. DIC and fluorescent images of the wild type and VPS8Dknockdown cells are shown in the left and right panels, respectively.
C: Severe mislocalization of Scr7p upon VPS8D knockdown. Growing cells expressing SCR7 -mNeon were fixed and the nuclei stained with DAPI. Fluorescent images of wild type and VPS8D knockdown cells are shown in the left and right halves, respectively, with focal planes selected from Z-stacks to illustrate the cell surface (left) and cell interior (right). Images were taken with a Zeiss Axio Observer 7 system.
Figure 4: