Fig. S4

Effects of cyp11a1 knockdown and pregnenolone treatment on wild-type and MZspg YCL microtubule dynamics

(a) The speed of microtubule growth at 40% epiboly plotted as EB3-mCherry track length (μm) over track duration (s) for stCoMo or MO1-cyp11a1 injected wild-type embryos. Morpholino injections were at 1-cell stage. Error bars indicate SEM (stCoMO n = 16; MO1-cyp11a1 n = 18).

(b) The microtubule growth speed was binned into speed intervals (bin size = 0.05 μm/s) at 40% epiboly for stCoMO or MO1-cyp11a1 injected wild-type embryos. Same dataset analyzed as in (a). Error bars indicate SEM.

(c) Microtubule organization of stCoMO (left column) or MO1-cyp11a1 (right column) injected MZspg embryos at indicated stages. Upper two rows show anti-ß-tubulin immunostaining. Lateral view, animal to the top, scale bar 100 μm. Bottom row shows YCL images of live embryos labeled with EMTB-3GFP. Scale bar 5 μm.

(d) The speed of microtubule growth at 40% epiboly plotted as EB3-mCherry track length (μm) versus track duration (s) for stCoMO or MO1-cyp11a1 injected MZspg embryos. Error bars indicate SEM (stCoMO n = 12; MO1-cyp11a1 n = 12).

(e) The speed of microtubule growth at 40% epiboly was binned into speed intervals (bin size = 0.05 μm/s) for stCoMO or MO1-cyp11a1 injected MZspg embryos. Error bars indicate SEM. Same dataset analyzed as in (f).

(f) Quantification of the speed of microtubule growth at sphere, 50% epiboly, and 70% epiboly plotted as EB3-mCherry track length (μm) over track duration (s) for control or pregnenolone (20 μM) treated wild-type embryos. Error bars indicate SEM (experimental controls: sphere: n = 11 embryos; 50% epiboly: n = 11; 70% epiboly: n = 8; pregnenolone treatment: sphere: n = 11, 50% epiboly n = 12; 70% epiboly: n = 8).

(g) The speed of microtubule growth at sphere, 50% epiboly, and 70% epiboly was binned into speed intervals (bin size = 0.05 μm/s) for control and pregnenolone treated wild-type embryos. Error bars indicate SEM. Same dataset analyzed as in (f).

(h) Average speed of microtubule growth of control or pregnenolone treated wild-type embryos, stages as indicated. Error bars - SEM. Same dataset analyzed as in (f). Data revealed no significant differences (Mann-Whitney test) of microtubule growth speed between control and pregnenolone treated embryos.