FIGURE 6

FIGURE 6

zfhx4 morphants resulted in defective cranial neural crest cell (CNCC) development and pharyngeal arch formation. (A–D″) Immunofluorescence images of the control and zfhx4 MO (E2I2)‐injected sox10:EGFP. Samples were stained with anti‐GFP antibody (NCCs), anti‐zfhx4 antibody, and 4′,6‐diamidino‐2‐phenylindole (DAPI, nucleus) at the 10‐somite stage (ss) and 24 hpf. (A–A″) zhfx4 protein demonstrated nuclear localization in the NCCs. Insets show a magnified view of the first pharyngeal arch (PA1, white dotted lines). Yellow arrowhead shows the co‐localization of the CNCCs with the zfhx4 protein. (B–B″) zfhx4 morphants showed decreased zfhx4 expression in the CNCCs. (C,D) zfhx4 morphants showed a decreased number of NCCs, leading to PA1 morphological defects. White dotted lines represent PA1 at 24 hpf. (C′–D″) Magnified view of PA1 in (C,D). (Control: N = 72, MO [E2I2]: N = 89). (E–F′) CNCC migration defect in zfhx4 morphants. zfhx4 MO was injected into sox10:EGFP embryos. Migrating CNCCs were visualized using sox10:EGFP at the 10 ss. The yellow arrowhead represents the position of the anteriormost midbrain and hindbrain, which are the starting sites for migrating CNCCs. The white arrowhead indicates the front edge of the migrating CNCCs. (Control: N = 55, MO [E2I2]: N = 38). e, eye; PA1, first pharyngeal arch. Scale bars: 100 μm in (C, D, E–F'); 50 μm in (A–B'', C'–D'').