Fig. 7

Genetic Interactions among mir92a, nog3, and Bmp Activity during Pharyngeal Cartilage Development

(A and B) 92a-MO-induced pharyngeal cartilage defects were rescued by coinjection of nog3-MO. Embryos injected with 4 ng 92a-MO alone or together with 0.1 ng nog3-MO at the one-cell stage, and harvested for Alcian blue staining at 80 hpf. Different degrees of rescue effects (NR0–NR2) were observed (A), and the ratio of each group was shown in (B).

(C) Ectopic expression of mir92a resulted in pharyngeal cartilage defects. Wild-type embryos were injected with 10 µM mir92a duplex at the one-cell stage, and stained at 80 hpf for cartilages with Alcian blue. Pharyngeal cartilage defects were categorized into groups P1–P3 (C), and the ratio of each group was indicated in the corresponding picture.

(D) mir92a duplex-induced pharyngeal cartilage defects were compromised by dorsomorphin treatment. Embryos injected with 10 µM mir92a duplex were treated in 5 µM dorsomorphin at 36 hpf and stained at 80 hpf for cartilage with Alcian blue. The percentage of embryos with different degrees of cartilage defects was calculated.

(E and F) mir92a duplex-induced apoptosis in the pharyngeal region. Tg(fli1:EGFP) transgenic embryos injected with 1 nl of 10 µM mir92a duplex were treated in 5 µM dorsomorphin at 36 hpf, subjected to TUNEL assay at 42 hpf, and observed by confocal microscopy. The representative images were lateral views of the first two pharyngeal arches (PA1 and PA2) (E). Scale bar, 20µm. The ratio of TUNEL- and GFP-positive cells (orange) to GFP-positive cells in the first and second pharyngeal arches were calculated from five embryos with one to two images each and presented in (F).

See also Fig. S7.