Phenotype evaluation of Alcian blue and immunofluorescence (IF) staining of the neuro- and viscero-cranium of various zebrafish larvae (zfl) groups. All zfl were at 4 dpf. N = 3 independent experiments per group. Scale bars, 100 μm. (A) overview of morphometric measurements of the upper and lower jaw structures of 4 dpf zfl: Ethmoid plate length (ep length) and palatoquadrate angle (pq angle) measurement lines presented in red in both whole-mount zfl head and separated viscero- and neuro-cranium. (B) box-plots of ep length and pq angle morphometric measurements. *P < 0.05, **P < 0.005 ***P < 0.0005. n, number of zfl. (C–H) representative examples of observed phenotypes, stained with Alcian blue (whole-mount heads) and IF (lower jaw structures): anti-DDR2; cartilage (anti-Col2a1). (C, D): Ddr2a Morpholino-knock-down (Ddr2a MO-KD) zfl, and controls (ctrl-MO); (E, F): ddr2a CRISPR knock-out (ddr2a CRISPR F0-KO) zfl and controls (scr-ctrl); (G, H): ddr2b KO zfl injected with ddr2a CRISPR, resulting in a ddr2a/ddr2b double-KO, and controls (scr-ctrl). Arrows indicate warped trabeculae; black brackets indicate a shortened ethmoid plate.
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