Figure 4. Loss of EBP-1/2/3 mildly... | Rockefeller University Press

Figure 4.

$Figure 4. Loss of EBP-1/2/3 mildly affects MT dynamics and does not perturb normal mitotic pulling force generation. (A) Quantification of anaphase centrosome rocking in WT and Δebp embryos. Maximum amplitudes are shown as means ± SEM. Unpaired Welch’s Student’s t test; *, P < 0.05. n = 10–23. (B) Example of spindle severing with a UV laser (purple) in a GFP::TBB-2 embryo. Arrows indicate the direction and relative speed of centrosome displacement upon bisection. (C) Quantification of centrosome velocities upon spindle severing in WT and Δebp embryos shown as means ± SEM. Unpaired Welch’s Student’s t test. n = 13–15. (D) Method for quantification of cortical MT residence time by TIRF microscopy of embryos expressing fluorescent tubulin. (E) Quantifications as illustrated in D for control and Δebp embryos. Bars represent means ± SEM. Mann–Whitney U test; ***, P < 0.001. 233 ≤ n ≤ 343 events from five embryos for each condition. Graphs show cumulative relative fractions of residence time distributions. Right graphs show density of cortical MT contacts as mean MTs/µm2 s−1 per embryo, and bars are means ± SEM. (F) Example images and quantifications of astral MT densities in control and Δebp mutant embryos, illustrated as radial intensity plots of fluorescent tubulin in metaphase (indicated in leftmost image). Plots were aligned and normalized to maximum values at the centrosome. n = 20 asters from 10 embryos per condition. Bars, 1 µm.$

Loss of EBP-1/2/3 mildly affects MT dynamics and does not perturb normal mitotic pulling force generation. (A) Quantification of anaphase centrosome rocking in WT and Δebp embryos. Maximum amplitudes are shown as means ± SEM. Unpaired Welch’s Student’s t test; *, P < 0.05. n = 10–23. (B) Example of spindle severing with a UV laser (purple) in a GFP::TBB-2 embryo. Arrows indicate the direction and relative speed of centrosome displacement upon bisection. (C) Quantification of centrosome velocities upon spindle severing in WT and Δebp embryos shown as means ± SEM. Unpaired Welch’s Student’s t test. n = 13–15. (D) Method for quantification of cortical MT residence time by TIRF microscopy of embryos expressing fluorescent tubulin. (E) Quantifications as illustrated in D for control and Δebp embryos. Bars represent means ± SEM. Mann–Whitney U test; ***, P < 0.001. 233 ≤ n ≤ 343 events from five embryos for each condition. Graphs show cumulative relative fractions of residence time distributions. Right graphs show density of cortical MT contacts as mean MTs/µm² s⁻¹ per embryo, and bars are means ± SEM. (F) Example images and quantifications of astral MT densities in control and Δebp mutant embryos, illustrated as radial intensity plots of fluorescent tubulin in metaphase (indicated in leftmost image). Plots were aligned and normalized to maximum values at the centrosome. n = 20 asters from 10 embryos per condition. Bars, 1 µm.