Live imaging of Dop1p reveals altered CVC morphology uponVPS8D knockdown.
A: Live imaging of the CVC contractile cycle in a wildtype cell expressing Dop1p-mNeon. The focal plane represents a cross-sectional view of the CVC, with 0.5 sec frame intervals. Seven successive images extracted from a video (Movie 3) are shown. Cells from growing cultures were immobilized using CyGEL, as described in Materials and Methods. Videos were captured using a Marianas spinning disc confocal microscope.
B: Live imaging of the CVC in VPS8D knockdown cell expressing Dop1p-mNeon. The cell selected contains two large bladders that undergo asynchronous contractions. Imaging conditions were the same as in Fig. 4A. The seven successive time-lapse images were extracted from Movie 4.
C: Live imaging of the CVC in VPS8D knockdown cell expressing Dop1p-mNeon. The cell selected contains multiple Dop1p-labeled vesicles dispersed in the cell posterior, concentrated in the region where the single stereotypical CVC bladder is typically located. Two of them contract over the course of the video. Imaging conditions were the same as in Fig. 4A, with 1 sec frame intervals. The six successive time-lapse images were extracted from Movie 5.
Figure 5: