Fig. S3

Quantification of migration and proliferation defects in miR-221 deficient and over-expressing embryos.

(A) Quantification of endothelial migratory speed in ISV sprouts in embryos injected with 10 ng of control MO or 10 ng miR-221 MO. “DA to 50 μM” is the initial sprouting from the dorsal aorta to the horizontal myoseptum, while “50 to 100 μM” is the migration from horizontal myoseptum to the DLAV. Measurements were made in 3 ISV sprouts each following time-lapse analysis from 2 embryos injected with control or miR- 221 Morpholino. * p<0.05.

(B) Confocal images of BrdU incorporation in Tg(kdrl:egfp)^la116 embryos at 27 hpf; lateral views. Embryos injected with 10 ng control MO, 10 ng miR-221 MO, or 500 μM miR-221 duplex. White arrowheads indicate stalk cells, arrows indicate DLAV or tip cells. Yellow arrowheads indicate BrdU-positive endothelial cells in the dorsal aorta and posterior cardinal vein in miR-221 MO-injected embryos. Scale bar is 50 μm.

(C) Quantification of BrdU-positive DLAV or endothelial tip cells within ISVs in Tg(kdrl:egfp)^la116 embryos 27 hpf injected with 10 ng of control, 10 ng miR-221 morpholino, 500 μM control duplex, or 500 μM miR-221 duplex. * p-value < 0.05.

(D) Quantification of BrdU-positive endothelial cells in posterior cardinal vein or dorsal aorta in Tg(kdrl:egfp)^la116 embryos at 27 hpf injected as in (D). In both (C) and (D) BrdU incorporation was determined for 4 adjacent ISVs per embryo in at least 8 embryos from 3 separate injections for each manipulation. The differences between paired samples in (D) are not statistically significant.