Supplementary Figure 5: In vivo localized loss and gain of function provokes the disruption of pinwheel-structures at the surface of the ventricle.

(a) Immunofluorescent detection of GFP in wholemounts from MT5-MMP infected mice. Note that infection with Lent-GFP-MT5-MMP also provokes the detachment of ependymocytes from the ventricle wall, however the effect is milder than with Lent-GFP-DN-CDH (Supplementary Fig. 4c). (b) Detection of S100β (blue), GFP (green) and EC-Ncadherin (red) in wholemounts of the infected ventricles. Note that the discrete membrane localization of EC-Ncadherin in GFP⁺ and non-infected (empty arrowheads) ependymal cells is disrupted in Lent-GFP-MT5-MMP infected cells (white arrowheads). Parallel control stainings can be seen in Supplementary Fig. 4b. (c–h) Wholemount staining with anti-GFAP (cyan), β-catenin and γ-tubulin (red) and GFP (green) to assess pinwheel structures (white dashed lines and arrows) in infected ventricles. (c,e,g: general view; d,f,h: detailed images). We were able to find some pinwheels in MT5-MMP-infected ventricles (white arrow), in contrast to DN-CDH infected areas. Scale bars: (a,b) 200 μm; 50 μm; 10 μm. (c,e,g) 10 μm. (d,f,h) 5 μm.