Fig. 4

Blocking miR-126 dependent spred1 suppression phenocopies miR-126 loss. (A) miR-126 binding site in the spred1 3′UTR. “Seed” sequence in bold, miR-126 in blue. The target protector morpholino (TP-MO) was designed to mask the “seed” sequence with its 3′ end to minimise binding to other miR-126 targets. (B) Injection of spred1 TP-MO phenocopies miR-126 loss. The formed TD can be visualized in uninjected control Tg(lyve1:DsRed2) larvae at 5 dpf. Injection of 800 pg of spred1 TP-MO reduced TD formation without affecting vISV number. At least 24 injected embryos analysed per experiment; 3 independent experiments. (C) flt4^+/- larvae are more sensitive to spred1 TP-MO injections. A significant reduction of TD formation can be observed in flt4^+/- but not in wild-type siblings injected with 400 pg of spred1 TP-MO. Injection of 80 pg of TP-MO did not affect TD development in either genotype. Minimum 8 embryos analysed per genotype per treatment group; 2 independent experiments. Error bars represent SEM. Unpaired Student's t-test was performed to test for statistical significance.