Fig. 4

IKNM Is Relatively Normal in the Absence of Stable Microtubules

(A) Polymerizing MT tips are stained by EB3. Centrin2 morphants show the same EB3-GFP kinetics as control embryos in Figure 1B. Dots at the end of lines mark the last position of EB3 comets followed.

(B) Antibody staining of α-tubulin (red) and acetylated tubulin (green) as in Figure 2C. Acetylated tubulin stains basal bodies (asterisks), but no stabilized MTs are observed spanning the epithelium (a). α-tubulin antibodies label mitotic spindles (arrow) and residual dynamic tubulin (b). A merge of (a) and (b) is shown in (c). Asterisks label stable basal bodies that are only stained by acetylated and not α-tubulin. DAPI (blue) counterstaining shows epithelial morphology (d).

(C) Images of Movie S6. Centrin2 morphants perform mitosis and cytokinesis at the apical membrane of the epithelium, after mitosis nuclei marked by H2B-RFP move away from this side. Centrosomes marked by γ-tubulin first serve as spindle pole bodies and then move back to apical side of epithelium (arrows).

(D) Images of Movies S7 and S8. Nuclei labeled by H2B-RFP move in apical and basal direction in centrin-2 morphants, indicated by white lines between false color red and yellow nuclei and the basal membrane of the epithelium.

(E) Nuclei still undergo rapid apical migration before mitosis as seen in the control cells.

(F) Stochastic motion velocity distributions for centrin morphant embryos, mosaic centrin morphant cells in control epithelium, and Colcemide-treated embryos.

(G) Stochastic motion MSD profile with linear fit (error bars show 95% confidence intervals) for centrin morphant embryos, mosaic centrin morphant cells in control epithelium, and Colcemide-treated embryos.

Scale bars represent 10 μm.