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Fig. 2
Dot1l Is a Direct Target of miR-216a
(A) Schematic of the reporter mRNA consisting of the coding sequence of GFP fused to the dot1l 3′ UTR. Three predicted miRNA recognition elements (MREs) are indicated. Predicted base pairing between MREs (shown in green) and the miR-216a sequence (shown in red).
(B) Embryos injected at the 1-cell stage with 100 pg of GFP-dot1l 3′ UTR, with or without 100 pg miR-216a, were examined for GFP expression at 1 day post fertilization (dpf). GFP expression was apparent in embryos injected with GFP-dot1l 3′ UTR, but it was reduced in embryos co-injected with miR-216a.
(C) Quantification of relative fluorescence in 1 dpf embryos injected with GFP reporter only or co-injected with miR-216a and the GFP reporter. Data represent means ± SEMs from 3 independent experiments. ∗∗p < 0.01 (Student’s t test).
(D) One-dpf-old embryos injected at the 1-cell stage with 100 pg of GFP-dot1l 3′ UTR carrying mutations in all miR-216a MREs. Embryos were injected with the mutant reporter, either alone or co-injected with miR-216a.
(E) Quantification of relative fluorescence in 1 dpf embryos injected with mutant GFP reporter alone or with co-injection of miR-216a. Data represent means ± SEMs from 3 independent experiments.
∗∗p < 0.01 (Student’s t test).
Scale bars, 500 μm.
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