Fig. 2

Apicobasal polarity loss and redistribution of primary cilia away from the ventricular surface in Afadin mutants. At E12.5, localization of the crumbs and Par complex-associated proteins (a, b) PALS1 and (c, d) PAR3 is similar in the control and mutant. e The apical domain protein Prominin-1 is localized adjacent to N-cadherin. In mutant, (f) localization of Prominin-1 is disrupted with elevated presence at lateral membranes of neuroepithelial cells lining the ventricle and in their cytoplasm. At E13.5, we observe disrupted localization of (g, h) PALS1 in the mutants, but not (i, j) PAR3. Prominin-1 localization, along with N-cadherin, is further disrupted in mutant (l) compared to control (k). m, n Lower (upper panels) and higher magnification images (lower panels) of the (m) E12.5 and (n) E13.5 dorsal telencephalon, visualizing the cilium-protein, Arl13b, in controls and mutants reveal the progressive loss of cilia from the ventricular surface in the mutant. (o, p) Quantifications of Arl13b puncta compared to Sytox Green-labeled nuclei (left panels) show that (o) total cilia number throughout the entire width of the telencephalon, as illustrated on the right panel, is not reduced in the mutant at E12.5 and E13.5, but decreases at E15.5. However, (p) the number of cilia within 20 μm of the ventricular surface, as illustrated on the right panel, is reduced at E12.5 and further decreased at E13.5 and E15.5 in mutant compared to control (mean ± s.e.m. Unpaired t-test; n = 3 embryos per group, 2 images per embryo). Scale bars: 10 μm (a-l); 20 μm (m and n, upper panels); 10 μm (m and n, lower panels)