Figure 2.
Figure 2. Klp2 constantly generates pulling forces, whereas dynein contribution increases during nuclear congression. (A, top) Overlay of differential interference contrast and fluorescence images of the SPB marker Sfi1-GFP shown for an individual zygote at 10 min before cell fusion (defined by cytoplasmic diffusion of mCherry-Atb2) until completion of nuclear congression at 26 min after cell fusion. Sfi1-GFP images were taken as 3D stacks at 1-min intervals. Dotted lines represent cell outlines. Dotted white line represents the line used to make the kymograph. (bottom) Kymograph representing Sfi1-GFP dynamics from −10 min until SPBs are juxtaposed. Yellow dotted lines indicate moment of cell fusion (t0). (B) SPB distance and velocity over time from individual example shown in A. Velocity obtained from a differential function of a second degree polynomial (black line) of SPB distances. (C) SPB distances over time normalized for each cell to mean SPB distance before cell fusion calculated from first 10 time points. Individual cells are depicted as light green trajectories (n = 25). Median is represented as a thick line, and upper and lower quartiles are shown as dotted lines. (D, left) Kymograph of Sfi1-GFP dynamics in an individual klp2Δ zygote. (right) SPB distances and velocity over time. Black line represents second degree polynomial function as approximation for SPB distance. (E) Sfi1-GFP dynamics in a subset of dhc1Δ zygotes (12/25 cells) depicted as kymograph (top) or as a function over time (bottom). Velocity obtained from a differential function of a linear regression (black line) of SPB distances. (F) Sfi1-GFP dynamics in a second subset of dhc1Δ zygotes (13/25 cells). Initial period without directed nuclear movement is highlighted. Velocity obtained from a differential function of a linear regression (black line) of SPB distances after initial period. (G and H) Median curves of normalized SPB distances (G) or velocities (H) over time in wild-type, klp2Δ, and a subset of dhc1Δ zygotes. Bars, 5 µm. n were collected from two independent experiments.

Klp2 constantly generates pulling forces, whereas dynein contribution increases during nuclear congression. (A, top) Overlay of differential interference contrast and fluorescence images of the SPB marker Sfi1-GFP shown for an individual zygote at 10 min before cell fusion (defined by cytoplasmic diffusion of mCherry-Atb2) until completion of nuclear congression at 26 min after cell fusion. Sfi1-GFP images were taken as 3D stacks at 1-min intervals. Dotted lines represent cell outlines. Dotted white line represents the line used to make the kymograph. (bottom) Kymograph representing Sfi1-GFP dynamics from −10 min until SPBs are juxtaposed. Yellow dotted lines indicate moment of cell fusion (t0). (B) SPB distance and velocity over time from individual example shown in A. Velocity obtained from a differential function of a second degree polynomial (black line) of SPB distances. (C) SPB distances over time normalized for each cell to mean SPB distance before cell fusion calculated from first 10 time points. Individual cells are depicted as light green trajectories (n = 25). Median is represented as a thick line, and upper and lower quartiles are shown as dotted lines. (D, left) Kymograph of Sfi1-GFP dynamics in an individual klp2Δ zygote. (right) SPB distances and velocity over time. Black line represents second degree polynomial function as approximation for SPB distance. (E) Sfi1-GFP dynamics in a subset of dhc1Δ zygotes (12/25 cells) depicted as kymograph (top) or as a function over time (bottom). Velocity obtained from a differential function of a linear regression (black line) of SPB distances. (F) Sfi1-GFP dynamics in a second subset of dhc1Δ zygotes (13/25 cells). Initial period without directed nuclear movement is highlighted. Velocity obtained from a differential function of a linear regression (black line) of SPB distances after initial period. (G and H) Median curves of normalized SPB distances (G) or velocities (H) over time in wild-type, klp2Δ, and a subset of dhc1Δ zygotes. Bars, 5 µm. n were collected from two independent experiments.

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