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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Dev Biol. 2021 Jan 20;472:98–114. doi: 10.1016/j.ydbio.2021.01.008

Figure 3. miR-31 directly suppresses Eve and Wnt1.

Figure 3

(A) Dual luciferase assays were conducted at mesenchyme blastula stage (24 hpf). The Rluc values were first normalized to the co-injected Firefly values. The ratios of normalized Rluc values with mutated miR-31 seed to the Rluc with wildtype (WT) miR-31 seed are presented. Luciferase readings of embryos injected with mutated (Mut) miR-31 3’UTR binding sites of Eve were increased significantly in comparison to embryos injected with the WT 3’ UTRs, indicating that miR-31 directly represses Eve. (B) Wnt1 has a mismatch of the first T nucleotide of the miR-31 seed sequence. Wnt1 is directly suppressed by miR-31. (C) Wnt5 contains a mismatch of the last C nucleotide of the miR-31 seed sequence. Wnt5 is not directly suppressed by miR-31 or experiences weak miRNA-mRNA binding affinity. 3–4 biological replicates. P-value was analyzed using Student’s t-test. All error bars represent SEM.