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Fig. 4 dmrt2b facilitates CNC cell proliferation and differentiation. (A,C,D) Representative confocal sections of pH3-positive cells in the first and second pharyngeal arches at the indicated stages. (B,E) Percentage of pH3-positive cells among GFP-positive CNC cells. n≥6 embryos for each condition. Significance was analyzed using unpaired t-tests. *P<0.05; **P<0.01; ***P<0.001. (F) Expression levels of col2a1 mRNA at 72 hpf. Red circles indicate the pharyngeal region. The left panels are lateral views with anterior to the left and the right panels are ventral views with the anterior to the top. (G,H) Immunostaining of Col2 protein in pharyngeal cartilages at 84 hpf. dmrt2b mutants (G) and Tg(fli1:EGFP) embryos injected with 4 ng dmrt2b MO (H) were stained with the indicated fluorescent antibodies. PA, pharyngeal arch; pq, palatoquadrate; ch, ceratohyal. Scale bars: 20 µm.