Figure 3. Redistribution of type 2 nodes... | Rockefeller University Press

Figure 3.

$Figure 3. Redistribution of type 2 nodes in cells during the cell cycle. (A) This time series of maximum intensity projections of reverse contrast fluorescence micrographs shows type 2 nodes marked with Blt1p-mEGFP emerging from the disassembling contractile ring and moving to the equator over 2 h. (B) Outcome plots of the time course of the localization of type 2 interphase nodes with time 0 at SPB separation, calculated as time from cell separation −80 min. Symbols: fraction of cells with ≥1 Blt1p node at new cell equator (closed squares); fraction of cells with nodes distributed along the cortex (defined as the first time point when the mean fluorescence at the old division site is less than or equal to the mean fluorescence at any other point along the length; closed triangles); and fraction of cells with Blt1p nodes maximally relocalized to the new cell equator (closed circles). (C) Histogram of the proportion of interphase nodes containing Blt1p only, Cdr2p only, or colocalized Blt1p and Cdr2p binned by stage of the cell cycle; same data as Fig. 1 E. (D) Plots of mean squared displacement (MSD) versus time step for five type 2 (Blt1p-mEGFP) nodes from a single interphase cell imaged at 2-s intervals. (E) Temporal color projections of node movements at 2-s intervals to compare the behavior of type 2 nodes marked with Blt1p-mEGFP (top) and type 1 nodes marked with Cdr2p-mEGFP at different stages of interphase (bottom). Each pair of micrographs has a reversed contrast sum fluorescence micrograph (three 300-nm z slices) on the left. Streaks of color show movements according to the time scale. White spots are stationary. The leftmost images are of one of two daughter cells still connected by a septum. Dotted ovals outline cells. (F) Plot of node diffusion coefficients in interphase cells as a function of the normalized distance from the new end of the cell showing that type 2 nodes diffuse faster near the cell tips than near the equator. Early interphase: cells still connected by septum or ≤8.5 µm in length; late interphase: cells >8.5 µm long. Type 2 Blt1p-mEGFP nodes (n = 49); type 2 Klp8p-mEGFP nodes (n = 23); type 1 Cdr2p-mEGFP nodes (n = 23). Bars, 2 µm.$

Redistribution of type 2 nodes in cells during the cell cycle. (A) This time series of maximum intensity projections of reverse contrast fluorescence micrographs shows type 2 nodes marked with Blt1p-mEGFP emerging from the disassembling contractile ring and moving to the equator over 2 h. (B) Outcome plots of the time course of the localization of type 2 interphase nodes with time 0 at SPB separation, calculated as time from cell separation −80 min. Symbols: fraction of cells with ≥1 Blt1p node at new cell equator (closed squares); fraction of cells with nodes distributed along the cortex (defined as the first time point when the mean fluorescence at the old division site is less than or equal to the mean fluorescence at any other point along the length; closed triangles); and fraction of cells with Blt1p nodes maximally relocalized to the new cell equator (closed circles). (C) Histogram of the proportion of interphase nodes containing Blt1p only, Cdr2p only, or colocalized Blt1p and Cdr2p binned by stage of the cell cycle; same data as Fig. 1 E. (D) Plots of mean squared displacement (MSD) versus time step for five type 2 (Blt1p-mEGFP) nodes from a single interphase cell imaged at 2-s intervals. (E) Temporal color projections of node movements at 2-s intervals to compare the behavior of type 2 nodes marked with Blt1p-mEGFP (top) and type 1 nodes marked with Cdr2p-mEGFP at different stages of interphase (bottom). Each pair of micrographs has a reversed contrast sum fluorescence micrograph (three 300-nm z slices) on the left. Streaks of color show movements according to the time scale. White spots are stationary. The leftmost images are of one of two daughter cells still connected by a septum. Dotted ovals outline cells. (F) Plot of node diffusion coefficients in interphase cells as a function of the normalized distance from the new end of the cell showing that type 2 nodes diffuse faster near the cell tips than near the equator. Early interphase: cells still connected by septum or ≤8.5 µm in length; late interphase: cells >8.5 µm long. Type 2 Blt1p-mEGFP nodes (n = 49); type 2 Klp8p-mEGFP nodes (n = 23); type 1 Cdr2p-mEGFP nodes (n = 23). Bars, 2 µm.