Fig. 3

Morphogenesis of the pharyngeal arches is disrupted in miR-27amorphants between 30 and 36hpf. (A) Live images of Tg(fli1a:eGFP)y1 embryos at 26, 30, 36, 48 hpf; lateral views with anterior to the left. Each pharyngeal arch is numbered. Embryos were injected with either 5 ng MO-ctl or MO-27 at the single cell stage and observed with confocal microscopy at the indicated stages. Time-lapse videos captured by spinning-disk confocal microscopy are shown in Supp. Movies 1 and 2. Scale bar, 100 µm. (B) Quantification of normalized and relative fli1a:eGFP fluorescence intensity in the pharyngeal arches of MO-ctl and MO-27 embryos at the indicated time points. The relative fluorescence intensities were normalized to the area of the arches selected and the fluorescence background in each image. Error bars indicate SEM and the number of embryos analyzed is indicated above the bars. For 26 hpf, n=3; for 30 hpf, n=5; for 36 hpf, n=4; for 48 hpf, n=4. Data represent three independent experimental trials. The n.s. non-significant, **p<0.01, ****p<0.0001 (Student's t-test). (C-D) Expression of the CNC marker dlx2a in embryos at 30 and 36 hpf. Embryos were injected with the corresponding morpholinos as described above. Pharyngeal arch expression domains are labeled. The indicated ratio represents the number of embryos with the represented phenotype/total number of observed embryos. Scale bar, 200 µm.