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Fig. 5 Metrns loss-of-function impairs Kupffer’s vesicle formation and function. (A) Confocal cross-section of 14 hpf wild type (WT) and triplMut counterstained with acetylated tubulin (AcTub) labeling the cilia of the Kupffer’s vesicle (KV). (B) The quantification of the cell number per KV in WT and triplMut embryos at 14 hpf shows a significant difference. (C) The KV diameter size measurement, (D) cilia number per KV quantification, and (E) individual cilia length measurement in WT and triplMut embryos at 14 hpf are all significantly decreased in triplMut (N WT: 23, N triplMut: 26, Student t-tests, ***p-value: 0.00028 for KV cell number, ***p-value: 9.7e-06 for KV diameter size, **p-value: 0.0012 for cilia number. For cilia length: average per KV and ***p-value: 5.8e-12). All error bars indicate standard deviation.(F) Single microbeads tracking in the KV of 12 hpf –14 hpf WT and triplMut embryos and the mean square displacement (MSD), revealing a directed trajectory in WT samples in contrast to triplMut, displaying short and undirected trajectories. Scale bar in (A): 20 µm.