Figure 2.
Figure 2. The formin For3 localization at the division site and its involvement in cytokinesis. (A) Timing (mean ± SD) of cytokinesis in for3Δ and wt at 23°C. *, P < 0.05; **, P < 0.005 compared with wt. This and subsequent p-values were determined using a two-tailed t test. Here and in later figures, ring assembly is from Rlc1 node appearance until a compact ring forms; ring maturation is from a compact ring to the start of ring constriction; ring constriction is from the start of constriction until the ring becomes a dot; and disassembly is from a dot until all the signal disappears from the division site. (B) For3 (green) localization to the division site. Arrowheads indicate cells with two SPBs (red) but no For3, whereas the arrow indicates a cell with two SPBs and For3. Graph (right) depicts 3D SPB distance in cells with short (<5 µm) spindles with (+) and without (−) For3 at the division site. (C–G) Cells were grown at 36°C for 2 h before imaging at 36°C. (C) For3 (green) localization in wt and cdc15-140 cells expressing CFP-Atb2 (red). Cells with similar spindle lengths are shown. Percentage of cells (with 2–7-µm spindles) with (wt) or without (cdc15-140) For3 at the division site is given. (D) For3 and Cdc15 localization in clumps in cdc12-112 mutants. (E) Quantification of clumps/nodes in (D) with Cdc15 only, For3 only, or colocalization of For3 and Cdc15. (F) Node condensation is impaired in cdc12-112 for3Δ double mutants compared with cdc12-112. (G) Quantification of clumps/nodes per cell in images as in F, subtracting two SPBs from the total. *, P < 0.0001 compared with cdc12-112. Box plots include 50% of the data inside the box, the line is the median, the whiskers indicate upper and lower bounds, and the circles outside the whiskers are outliers. (H) Timing of contractile ring stages (mean ± SD) in indicated Rlc1-3GFP labeled strains grown at 30°C for 2 h before imaging at 30°C. *, P < 0.05; **, P < 0.005; ***, P < 0.0005 in t tests compared with wt (except where the bracket indicates). (I) Staining of actin filaments in wt, cdc12-112, and cdc12-112 for3Δ cells. Cells were fixed at 36°C after growing at 36°C for 2 h. Single planes near the top of the cells and maximum intensity projections are shown. The meshwork/ring of filaments at the division site is marked (arrowheads). (J) Quantification of cells as in I. Percent cells with actin meshwork and rings (wt only) at the division site. Bars, 5 µm.

The formin For3 localization at the division site and its involvement in cytokinesis. (A) Timing (mean ± SD) of cytokinesis in for3Δ and wt at 23°C. *, P < 0.05; **, P < 0.005 compared with wt. This and subsequent p-values were determined using a two-tailed t test. Here and in later figures, ring assembly is from Rlc1 node appearance until a compact ring forms; ring maturation is from a compact ring to the start of ring constriction; ring constriction is from the start of constriction until the ring becomes a dot; and disassembly is from a dot until all the signal disappears from the division site. (B) For3 (green) localization to the division site. Arrowheads indicate cells with two SPBs (red) but no For3, whereas the arrow indicates a cell with two SPBs and For3. Graph (right) depicts 3D SPB distance in cells with short (<5 µm) spindles with (+) and without (−) For3 at the division site. (C–G) Cells were grown at 36°C for 2 h before imaging at 36°C. (C) For3 (green) localization in wt and cdc15-140 cells expressing CFP-Atb2 (red). Cells with similar spindle lengths are shown. Percentage of cells (with 2–7-µm spindles) with (wt) or without (cdc15-140) For3 at the division site is given. (D) For3 and Cdc15 localization in clumps in cdc12-112 mutants. (E) Quantification of clumps/nodes in (D) with Cdc15 only, For3 only, or colocalization of For3 and Cdc15. (F) Node condensation is impaired in cdc12-112 for3Δ double mutants compared with cdc12-112. (G) Quantification of clumps/nodes per cell in images as in F, subtracting two SPBs from the total. *, P < 0.0001 compared with cdc12-112. Box plots include 50% of the data inside the box, the line is the median, the whiskers indicate upper and lower bounds, and the circles outside the whiskers are outliers. (H) Timing of contractile ring stages (mean ± SD) in indicated Rlc1-3GFP labeled strains grown at 30°C for 2 h before imaging at 30°C. *, P < 0.05; **, P < 0.005; ***, P < 0.0005 in t tests compared with wt (except where the bracket indicates). (I) Staining of actin filaments in wt, cdc12-112, and cdc12-112 for3Δ cells. Cells were fixed at 36°C after growing at 36°C for 2 h. Single planes near the top of the cells and maximum intensity projections are shown. The meshwork/ring of filaments at the division site is marked (arrowheads). (J) Quantification of cells as in I. Percent cells with actin meshwork and rings (wt only) at the division site. Bars, 5 µm.

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