Fig. 5

Ttc28 Is Required to Regulate Microtubule Dynamics

(A) Schematic of ttc28 single guide RNA targeting site and the sequences of ttc28^stl362 and ttc28^stl363 mutant alleles. Asterisk indicates premature stop codon.

(B) qPCR analyses of ttc28 transcript levels in WT, MZttc28^{stl362/stl362}, and MZttc28^{stl363/stl363} mutants at two-cell stage. Two different sets of primers were used and normalized to β-actin. ^∗∗∗∗p < 0.0001, Student’s t test. Error bars represent SD. N = 3 biological repeats.

(C) Representative time-lapse still images of YCL microtubules at 1.75–2.5 hpf in WT, MZttc28^{stl363/stl363}, and HA-Ttc28-mCherry RNA-injected (900 pg) MZttc28^{stl363/stl363}Tg[ef1α:dclk-GFP] embryos. Scale bar, 30 μm.

(D) Quantification of YCL microtubule density from time-lapse videos in WT, MZttc28^{stl363/stl363}, and HA-Ttc28 or HA-Ttc28-mCherry RNA-injected (900 pg) MZttc28^{stl363/stl363} embryos at 1.75–2.5 hpf. Error bars represent SEM. N, number of embryos.

(E) Representative time-lapse still images of microtubule dynamics during cytokinesis in WT and MZttc28^{stl363/stl363} embryos using Tg[ef1α:dclk-GFP]. Scale bar, 30 μm.

(F) Quantification of midzone microtubule assembly rates in WT and MZttc28^{stl363/stl363} blastulae at 1.75–2.5 hpf. ^∗∗∗∗p < 0.0001, paired Student's two-tailed t test. Error bars represent SEM. N, number of embryos.

See also Figures S5 and S6, Videos S6 and S7.