Figure 4: Using morpholino target protector (TP) to show that miR-1 represses zebrafish sars-3′UTR. | Nature Communications

Figure 4: Using morpholino target protector (TP) to show that miR-1 represses zebrafish sars-3′UTR.

From: miR-1 and miR-206 target different genes to have opposing roles during angiogenesis in zebrafish embryos

Figure 4

(a) Sequence of the target site, cognate sars-TP-MO (sars-TP), and miR-1 were indicated. The full length of sars-3′UTR was inserted downstream of an EGFP expression vector. (b) In the control group, the GFP signal was observed throughout the embryos injected with egfp-mRNA plus control-MO. (c) The GFP signal in truck somites, shown at right lower corner, was greatly reduced in embryos injected with control-MO plus egfp-mRNA containing WT sars-3′UTR. (d) The GFP signal in truck somite was restored when the miR-1 target site on sars-3′UTR was mutated. (e) The GFP signal in truck somite was also restored in embryos injected with EGFP-WT sars-3′UTR plus sars-TP-MO. (f) The reduced GFP caused by EGFP-sars-3′UTR could be restored by coinjection of miR-1-MO1. (g) The reduced GFP caused by EGFP-sars-3′UTR could not be restored by coinjection of miR-206-MO. (h) Western blot analysis to detect the proteins of SARS and VegfAa in the WT, control-MO-injected and sars-TP-MO-injected embryos at 48 hpf. (i) The statistical analyses of the average relative intensities of SARS and VegfAa performed on densitometry from WT, control-MO-injected and sars-TP-MO-injected embryos after three independent experiments. Data are presented as mean±s.d. (n=3). Student's t-test was used to determine significant differences between each construct and control within each group (***P<0.005). GAPDH served as an internal control. (j–q) Embryos derived from line Tg(fli1:EGFP) were injected with materials as indicated, and ISV patterns were observed at 28 and 48 hpf. Scale bar: (b–g) 300 μM; (j–q) 100 μM.

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